Serodiagnosis of Borreliosis: Indirect Immunofluorescence Assay, Enzyme-Linked Immunosorbent Assay and Immunoblotting
Iwona Wojciechowska-Koszko • Iwona Mączyńska • Zbigniew Szych • Stefania Giedrys-Kalemba
Abstract Lyme disease is an infectious, multi-system, tick-borne disease caused by genospecies of Borrelia burgdorferi bacteria sensu lato, characterized by remarkable heterogeneity. In this situation choosing an optimal antigen array for diagnostic tests seems problematic. The serological tests for borrelia routinely done in laboratories often produce ambiguous results, which makes a proper diagnosis rather complicated and thus delays the implementation of an appropriate treatment regimen. Thirtyseven outpatients and eight inpatients with suspected borreliosis diagnosis hospitalized at the Clinics of the Pomeranian Medical University (Szczecin, Poland), participated in the study. In order to detect the antibodies against Borrelia sensu lato three kinds of serological tests were used: indirect immunofluorescence assay (IIFA), enzyme-linked immunosorbent assay (ELISA), and immunoblot. The IIFA and immunoblot tests conducted on 45 patients (100%) produced positive results for both the IgM and IgG antibody types. In the case of ELISA, positive or borderline results were observed in only 24 patients (53.3%). The immunoblot test for IgM most frequently detected antibodies against the outer surface protein C (OspC) antigen (p25), and, in the case of IgG, against the recombinant variable surface antigen (VlsE). The IIFA screening test used for diagnosing Lyme borreliosis produced the highest percentage of positive results, which were then confirmed by immunoblot, but not by ELISA. Therefore using only ELISA as a screening test or for diagnosing Lyme borreliosis seems debatable.
Keywords Lyme borreliosis ELISA IIFA Immunoblot VlsE protein OspC protein
Relapse of Acute Lymphoblastic Leukemia in Children in the Context of Microarray Analyses
Joanna Szczepanek • Jan Styczyński • Olga Haus • Andrzej Tretyn • Mariusz Wysocki
Abstract Over the last four decades the treatment of patients with newly diagnosed childhood acute lymphoblastic leukemia (ALL) has improved remarkably. However, still about 20% of children with ALL relapse despite risk-adapted polychemotherapy. The prognosis of relapsed ALL is relatively poor, even with modern aggressive chemotherapy. Identification of the biological and genetic mechanisms contributing to recurrence in patients with ALL is critical for the development of effective therapeutic strategies to treat refractory leukemic patients. Allogeneic hematopoietic stem-cell transplantation is the treatment of choice for many children with relapsed ALL. The gene expression profile obtained by microarray technology could provide important determinants of the drug response and clinical outcome in childhood ALL. Incorporation of the data on expression levels of newly identified genes into existing strategies of risk stratification might improve clinical management. Current microarray data show correlation of in vitro drug resistance with significant patterns of gene expression and explain clinical differences between early and late relapse. Genes involved in cell proliferation, self-renewal and differentiation, protein biosynthesis, carbohydrate metabolism, and DNA replication and repair are usually among those highly expressed in relapsed lymphoblasts. Current status and future perspectives of microarray data on gene expression and drug resistance profile in relapsed pediatric ALL are discussed in this review.
Keywords Gene expression profile Copy number abnormalities Microarray Children Acute lymphoblastic leukemia Relapse
CD46 Plasticity and Its Inflammatory Bias in Multiple Sclerosis
Siobhan Ni Choileain • Anne L. Astier
Received: 7 July 2010 / Accepted: 16 September 2010 / Published online: 26 January 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Known as a link to the adaptive immune sys-
tem, a complement regulator, a ‘‘pathogen magnet’’ and
more recently as an inducer of autophagy, CD46 is the
human receptor that refuses to be put in a box. This review
summarizes the current roles of CD46 during immune
responses and highlights the role of CD46 as both a pro-
moter and attenuator of the immune response. In patients
with multiple sclerosis (MS), CD46 responses are over-
whelmingly pro-inflammatory with notable defects in
cytokine and chemokine production. Understanding the
role of CD46 as an inflammatory regulator is a distant goal
considering the darkness in which its regulatory mecha-
nisms reside. Further research into the regulation of CD46
expression through its internalization and processing will
undoubtedly extend our knowledge of how the balance is
tipped in favor of inflammation in MS patients.
Keywords CD46 Multiple sclerosis T cell
Regulation
Tuning of CD40–CD154 Interactions in Human B-Lymphocyte
Activation: A Broad Array of In Vitro Models for a Complex
In Vivo Situation
Sonia Ne´ron • Philippe J. Nadeau •
Andre´ Darveau • Jean-Franc¸ois Leblanc
Received: 30 March 2010 / Accepted: 19 August 2010 / Published online: 14 January 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Naive and memory B-lymphocyte populations
can be activated through the binding of CD154 to CD40, a
receptor that is constitutively expressed on the surface of
these cells. Models based on the in vitro stimulation of
human B lymphocytes through CD40 have greatly con-
tributed to our understanding of the human immune
response in healthy individuals and patients suffering from
immune disorders. The nature of the engineered CD40
ligands is as diverse as the in vitro models used in studies
of CD40-activated B lymphocytes. Monoclonal anti-CD40
antibodies, recombinant CD154 proteins, soluble CD154?
membranes as well as CD154? cell lines have turned out to
be very useful tools, and are still in use today. As for any
receptor–ligand interaction, parameters such as duration
and strength of contact, timing, affinity, and receptor
density are major determinants of CD40 binding by CD154
or anti-CD40. Furthermore, variation in the intensity of
CD40 stimulation has been shown to influence prolifera-
tion, differentiation and immunoglobulin secretion of
human hybridomas, B-cell lines, tonsil and blood B lym-
phocytes. The objective of this review is to present an
overview of the great diversity of CD40 agonists used in in
vitro models of B-lymphocyte activation, with a particular
emphasis on variations in the resulting strength of
CD40 signaling generated by these models. A better
understanding of these models could open up new avenues
for the rational use of human B lymphocytes as antigen-
presenting cells in cellular therapies.
Keywords CD40–CD154 intensity B lymphocytes
In vitro models
Antiviral Signaling Through Retinoic Acid-Inducible
Gene-I-Like Receptors
Tomoh Matsumiya • Tadaatsu Imaizumi •
Hidemi Yoshida • Kei Satoh
Received: 16 July 2010 / Accepted: 20 August 2010 / Published online: 14 January 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The innate immune system is essential for the
first line of host defense against micropathogens. In virus-
infected cells, exposed viral nucleotides are sensed by
pattern recognition receptors (PRRs), resulting in the
induction of type I interferon. Retinoic acid-inducible
gene-I-like receptors (RLRs) are a member of PRRs and
are known to be crucial molecules in innate immune
responses. Upon viral recognition, RLRs recruit their spe-
cific adaptor molecules, leading to the activation of
antiviral signaling molecules including interferon regula-
tory factor-3 and nuclear factor-jB. Mitochondrial antiviral
signaling (MAVS) protein is also known as one of the
adaptor molecules responsible for antiviral signaling trig-
gered by RLRs. Recent reports have identified numerous
intracellular molecules involved in the antiviral responses
mediated by RLRs/MAVS. Several viral proteins interfere
with the RLR/MAVS signaling, allowing the virus to evade
the host defense. In this review, we comprehensively
update RLR-dependent antiviral signaling with special
reference to the RLRs/MAVS-mediated responses.
Keywords Innate immunity RLRs MAVS
Dual Peripheral Actions of Immune Cells in Neuropathic Pain
Halina Machelska
Received: 16 March 2010 / Accepted: 13 September 2010 / Published online: 14 January 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Ability to perceive physiological pain is
essential in protecting the individual from tissue destruc-
tion. In contrast, pathological chronic pain is an expression
of maladaptive alterations outlasting its biological useful-
ness. In such conditions even eating, speaking or wearing
clothes might be painful, as in neuropathic pain. Such pain
is caused by diseases or injuries affecting nerves (e.g.
diabetes, trigeminal neuralgia or amputation). Neuropathic
pain is not an exclusive neuronal phenomenon but also
involves immune responses. Damaged peripheral nerves
are infiltrated by mast cells, granulocytes, macrophages
and T lymphocytes. It is widely emphasized that these
cells, via secretion of inflammatory mediators (e.g. proin-
flammatory cytokines, chemokines), contribute to the
generation of neuropathic pain. However, leukocytes are
also a source of analgesic mediators such as anti-inflam-
matory cytokines and opioid peptides. Recent findings
indicate that immune cell-derived opioid peptides can
interact with opioid receptors in the injured nerves and
ameliorate neuropathic pain. Targeting opioid-containing
immune cells might represent a new disease-modifying
approach based on the use of beneficial effects of neuro-
inflammation in painful neuropathies. This review analyzes
both detrimental and advantageous actions of leukocytes at
peripheral nerves in neuropathic pain.
Keywords Analgesia Cytokines Inflammation
Leukocytes Nerve injury Opioids
Legal and Ethical Aspects of Ghostwriting in Medicine
Agata Wnukiewicz-Kozłowska
Received: 15 July 2010 / Accepted: 6 August 2010 / Published online: 14 January 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Ghostwriting in medicine represents a highly
significant problem in both legal and ethical terms, and
particularly in recent years owing to a sudden increase in
the practice. The main goal in this paper is to present the
legal and ethical rules connected with the ghostwriting
phenomenon. The problem is presented in the context of
international, European and Polish law. From a legal per-
spective the issues bound up with ghostwriting are those of
the notion and authorship of a work, authors’ rights (both
moral rights and copyright), the content of these rights and
their nature, as well as the question of the transfer of such
rights. From the point of view of ethics there arises a need
for reflection on honesty, accuracy and credibility as
defined both generally and scientifically.
Keywords Ghostwriting Authors’ rights
(copyright and moral rights) Transfer of authors’ rights
Validation of Basophil CD164 Upregulation
for Pollen Allergy Diagnosis
Anna Wolanczyk-Medrala • Wojciech Barg • Jerzy Liebhart •
Bernard Panaszek • Grazyna Nadobna • Marta Litwa •
Grzegorz Gogolewski • Wojciech Medrala
Received: 26 February 2010 / Accepted: 23 July 2010 / Published online: 28 September 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract The aim of the study was to evaluate the sensi-
tivity and specificity of allergen-induced basophil CD164
upregulation in patients with seasonal allergic rhinitis
caused by allergy to grass pollens. This study was performed
in 24 patients with allergy to grass pollens, and in 25 healthy
controls. The protocol for allergen-induced basophil CD164
upregulation consisted of whole blood samples processing
and staining with anti-CCR3/anti-CD164 antibodies added
to a buffer at the beginning of stimulation. We observed
dose-dependent allergen-induced basophil CD164 upregu-
lation with 100% of specificity in both used allergen
concentrations (12 and 1.2 ng/ml). Higher allergen
concentration resulted in 100% and lower concentration in
only 70.83% sensitivity. We have observed in the patients
statistically significant correlations between anti-IgE
stimulation and both allergen concentrations (for 12 ng/ml,
r = 0.71, p \ 0.0001; and for 1.2 ng/ml, r = 0.64,
p \ 0.001). We conclude that assessment of allergen-
induced basophil CD164 upregulation is a very useful
method for in vitro determination of allergy to grass pollens.
This method seems to be a very promising tool in laboratory
testing of allergies to other allergens.
Keywords Allergy Basophil CD164
Flow cytometry Basophil activation test
How do Tumors Actively Escape from Host Immunosurveillance?
Jacek R. Wilczyn´ ski • Markus Duechler
Received: 1 March 2010 / Accepted: 28 May 2010 / Published online: 5 October 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract The immunological background for the process
of tumor growth is still obscure. However, our understanding
of what happens could have important consequences,
namely in the context of cancer immunotherapy. A tumor is
able to grow in the host environment either because it is
recognizable as normal tissue and tolerated by host immune
cells, or because it can ‘‘escape’’ from host immunosur-
veillance. According to the second option the mechanisms of
tumor recognition and consequent destruction are actively
disturbed by such processes as: change of tumor immuno-
genicity, production of tumor-derived regulatory molecules,
and interaction of cancer cells with tumor-infiltrating
immune cells. The results of studies devoted to the problem
of immunoregulation in the tumor environment seem to
support the ‘‘escape’’ hypothesis.
Keywords Tumor Immunosurveillance
Immunoediting
The Center of Accessibility: Db Control of V(D)J Recombination
Michael L. Sikes • Ruth E. McMillan •
Justin M. Bradshaw
Received: 14 June 2010 / Accepted: 23 July 2010 / Published online: 2 October 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Developmental patterning of antigen receptor
gene assembly in lymphocyte precursors correlates with
decondensation of the chromatin surrounding individual
gene segments. Ongoing V(D)J recombination is associ-
ated with hyperacetylation of histones H3 and H4 and the
expression of sterile germline transcripts across the region
of recombinational accessibility. Likewise, histone acetyl-
transferase and SWI/SNF chromatin remodeling complexes
each appear to be required for recombination, and the
PHD-finger of RAG-2 preferentially associates with
recombination signal sequence (RSS) chromatin that con-
tains H3 trimethylated on lysine 4. However, the regulatory
mechanisms that direct chromatin alteration and rear-
rangement have proven elusive, due in large part to the
interdependency of individual stages in gene activation, our
limited understanding of functional significance of changes
to the histone code, and the difficulty of modeling
recombinational accessibility in existing experimental
systems. Examining Tcrb assembly in developing thymo-
cytes, we review the central roles of RSS elements and
germline promoters as foci for epigenetic reorganization of
recombinationally accessible gene segments in light of
recent findings and persistent questions.
Keywords T cell receptor Tcrb Transcription
V(D)J recombination Thymocytes Double negative
Dysregulation of Thymic Clonal Deletion and the Escape
of Autoreactive T Cells
Charles J. Kroger • Rafael R. Flores •
Maurice Morillon • Bo Wang • Roland Tisch
Received: 17 May 2010 / Accepted: 25 June 2010 / Published online: 26 September 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Events ongoing in the thymus are critical for
deleting developing thymocytes specific for tissue antigens,
and establishing self-tolerance within the T cell compart-
ment. Aberrant thymic negative selection, however, is
believed to generate a repertoire with increased self-reac-
tivity, which in turn can contribute to the development of T
cell-mediated autoimmunity. In this review, mechanisms
that regulate the efficacy of negative selection and influ-
ence the deletion of autoreactive thymocytes will be
discussed.
Keywords Antigen-presenting cells Autoimmunity
Central tolerance Thymocytes
Flaws in the Medical Literature: Who’s to Blame?
Jerome P. Kassirer
Interleukin-27: Biological Properties and Clinical Application
Marek Jankowski • Piotr Kopin´ ski •
Anna Goc
Received: 22 October 2009 / Accepted: 26 February 2010 / Published online: 26 September 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Interleukin (IL)-27 is a novel cytokine secreted
by stimulated antigen-presenting cells. Initial studies on the
biology of IL-27 provided evidence for its role in the initi-
ation of TH1 responses; however, subsequent work has
indicated that IL-27 has broad inhibitory effects on TH1,
TH2, and TH17 subsets of T cells as well as the expansion of
inducible regulatory T cells. The involvement of IL-27 in the
regulation of angiogenesis and antiviral response has also
recently been reported. The aim of this review is to highlight
the potential areas of IL-27 clinical application, especially
the management of neoplastic and viral diseases as well as
autoimmune disorders, including rheumatoid arthritis and
multiple sclerosis. The review will also serve to elaborate on
the molecular mechanisms involved in the expression of this
cytokine and signaling from the IL-27 receptor.
Keywords IL-27 Cancer Multiple sclerosis
Rheumatoid arthritis HIV Immunotherapy
Tumor-Resident CD8+ T-cell: The Critical Catalyst
in IL-12-Mediated Reversal of Tumor Immune Suppression
Nejat K. Egilmez • Mehmet O. Kilinc
Received: 6 April 2010 / Accepted: 21 June 2010 / Published online: 26 September 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Tumor-resident T cells display a functionally
impaired effector/memory (Tem) phenotype. Sustained
intratumoral administration of IL-12, on the other hand,
can restore cytolytic function to pre-existing CD8? Tem,
resulting in effective tumor kill. Whereas cytotoxic T
lymphocytes (CTL) are generally assumed to mediate
tumor regression via direct tumor cytotoxicity, recent work
revealed that activated CD8? Tem mobilize a systemic,
multi-component effector cascade that includes both innate
and adaptive immune mechanisms. Here we summarize
these mechanisms, review how tumor-resident CD8? Tem
orchestrate this cascade and discuss the potential clinical
implications of these findings.
Keywords Tumor microenvironment Immune therapy
CD8? T cells Immune suppression IL-12
Detection of b-Herpesviruses in Polish Adult Cord Blood Stem
Cell Recipients by Real-Time PCR: Single Centre Study
Tomasz Dziecia˛tkowski • Maciej Przybylski • Grzegorz Władysław Basak •
Tigran Torosian • Agnieszka Tomaszewska • Wiesław Wiktor Je˛drzejczak •
Gra_zyna Młynarczyk
Received: 16 November 2009 / Accepted: 1 March 2010 / Published online: 25 September 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Umbilical cord blood transplantation (UCBT)
is known to be associated with increased risk of infections,
compared to bone marrow or peripheral blood stem cell
transplantation. In viral diseases for which specific treat-
ment is available, real-time PCR assays are reliable
diagnostic tools for timely initiation of appropriate therapy
and for rapid assessment of the efficacy of antiviral treat-
ment strategies. A retrospective review of samples from a
group of seven adult cord blood stem cell recipients was
made. Serum samples taken up to 180 days after trans-
plantation were examined with quantitative real-time PCR
for measurement of viral load (CMV, HHV-6, and HHV-7).
Cytomegalovirus (CMV) DNA was detected in samples
taken from four patients (57%) in the period of 20–80 days
after transplantation. Products of amplification of human
herpesvirus 6 (HHV-6) DNA were found in samples taken
between days 25 and 37 following UCBT from only one
patient (14%). On the other hand, the majority of patients
(n = 6, 86%) had HHV-7 DNA detected in the period
15–58 days after transplantation. Co-infection with HHV-7
was demonstrated at onset of all episodes of microbiolog-
ically confirmed CMV or HHV-6 infection. Our obser-
vations indicate that real-time PCR is not only useful for
monitoring herpesviral infections in transplant recipients,
but is also a powerful method for clarifying the relation-
ships between the viral load and clinical symptoms. Further
investigation with a much larger group of patients will be
needed to confirm these observations and translate them
into a clinical approach.
Keywords Umbilical cord blood transplantation
Post-transplant infections Herpesviruses Real-time PCR
Iron and Immunity: Immunological Consequences
of Iron Deficiency and Overload
Bobby J. Cherayil
Received: 5 May 2010 / Accepted: 23 July 2010 / Published online: 28 September 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract The influence of iron on immune function has
been long appreciated. However, the molecular basis for
this interaction is less well understood. Recently, there
have been several important advances that have shed light
on the mechanisms that regulate mammalian iron metab-
olism. The new insights provide a conceptual framework
for understanding and manipulating the cross-talk between
iron homeostasis and the immune system. This article will
review what is currently known about how disturbances of
iron metabolism can affect immunity and how activation of
the immune system can lead to alterations in iron balance.
Keywords Iron Metabolism Macrophage
Inflammation Lymphocyte
Autoimmune Versus Oligodendrogliopathy: The Pathogenesis
of Multiple Sclerosis
Jin Nakahara • Sadakazu Aiso • Norihiro Suzuki
Received: 2 February 2010 / Accepted: 6 April 2010 / Published online: 31 July 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Multiple sclerosis (MS) is the most common
inflammatory demyelinating disease of the central nervous
system (CNS); it affect millions of patients worldwide and
the number of patients is on the rise. Current treatment
options are fairly limited and there is a strong unmet need
for disease-targeted therapies for MS. The most widely
accepted hypothesis for the pathogenesis of MS is that it is
a primary autoimmune disease in which myelin-specific T
cells play a central role in the progression of demyelina-
tion. According to this hypothesis, a powerful immune
suppression or a reconstruction of the immune system to
abrogate disease-specific leukocytes early in the develop-
ment of the disease is expected to halt or even reverse the
disease, since remyelination is an exceptionally efficient
regenerative process in the CNS. However, recent neuro-
pathological studies have provided evidence of primary
oligodendrogliopathy as a cause of demyelination, sug-
gesting that immune reactions may be a mere secondary
event in the course of MS. On the other hand, some recent
clinical trial results of new immune-suppressive treatments
showed a nearly complete blockade of relapses and
significant, albeit incomplete, neurological improvement.
Therefore, which hypothesis—autoimmunity or oligoden-
drogliopathy—lights the correct path to a ‘‘cure’’ for MS?
Keywords Multiple sclerosis Pathogenesis
Autoimmune Oligodendrogliopathy
Monocyte and Macrophage Abnormalities in Systemic Lupus
Erythematosus
Yi Li • Pui Y. Lee • Westley H. Reeves
Received: 1 January 2010 / Accepted: 9 April 2010 / Published online: 31 July 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Systemic lupus erythematosus (SLE) is a
chronic autoimmune disease with profound effects on
multiple organ systems. In patients with SLE, the immune
system is subverted to target numerous self antigens and
the ensuing inflammatory response elicits a vicious cycle of
immune-cell activation and tissue damage. Both genetic
and environmental factors are essential for the develop-
ment of this debilitating condition, although the exact
cause remains unclear. Early studies on the pathogenesis of
lupus centered on the adaptive immune system as lym-
phocyte abnormalities were thought to be the primary
cause of autoimmunity. In the past decade, however, this
paradigm has shifted with rapid advances in the field of
innate immunity. These developments have yielded
important insights into how the autoimmune response in
SLE is initiated and maintained. Monocytes and macro-
phages are an essential arm of the innate immune system
with a multitude of immunological functions, including
antigen presentation, phagocytosis, and cytokine produc-
tion. Aberrations of monocyte/macrophage phenotype and
function are increasingly recognized in SLE and animal
models of the disease. In this review we summarize the
current knowledge of monocyte/macrophage abnormalities
in human SLE and discuss their implications for under-
standing the pathogenesis of lupus.
Keywords Systemic lupus erythematosus Monocytes
Macrophages
Polymorphisms in the Genes Encoding TGF-b1, TNF-a,
and IL-6 Show Association with Type 1 Diabetes Mellitus
in the Slovak Population
Juraj Javor • Stanislav Ferencik • Maria Bucova •
Martina Stuchlikova • Emil Martinka • Lubomir Barak •
Lujza Strbova • Hans Grosse-Wilde • Milan Buc
Received: 3 December 2009 / Accepted: 19 March 2010 / Published online: 5 August 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Numerous cytokines have been shown to par-
ticipate in the pathogenesis of type 1 diabetes (T1D). As gene
polymorphisms can influence cytokine production or
function, they may potentially contribute to genetic predis-
position to the disease. The aim of this study was therefore to
investigate the role of 22 single nucleotide polymorphisms
(SNPs) in 13 cytokine and cytokine receptor genes in genetic
susceptibility to T1D. Polymerase chain reaction with
sequence-specific primers was used to genotype cytokine
SNPs and HLA-DRB1 alleles in 151 diabetics and 140
healthy individuals of Slovak origin. Univariate analysis
showed that transforming growth factor (TGF)-b1 codon
10 TT homozygotes were significantly more susceptible to
developing T1D than C allele carriers (Pc = 0.0066, OR =
2.46). Furthermore, tumor necrosis factor (TNF)-a –308
A allele carriers were also significantly overrepresented
among the diabetics (Pc = 0.0031, OR = 2.62); however,
the association of the –308 A allele with T1D might be due
to its strong linkage disequilibrium with the susceptibility
allele HLA-DRB1*0301. An association was also found
with interleukin (IL)-6 –174 G/C and nt565 G/A SNPs;
however, its significance was lost when statistical correction
was applied. These data suggest that the TGF-b1 codon 10
SNP is among numerous genetic variations with small
individual effects on T1D development. Moreover, a possi-
ble role of TNF-a and IL-6 SNPs cannot be ruled out,
although their association with T1D was due to strong LD
with the HLA class II susceptibility allele or did not with-
stand statistical correction, respectively.
Keywords Cytokines Interleukin-6 Single nucleotide
polymorphisms Transforming growth factor-b1
Tumor necrosis factor-a Type 1 diabetes
Genetic Modification of T Cells Improves the Effectiveness
of Adoptive Tumor Immunotherapy
Marek Jako´bisiak • Jakub Goła˛b
Received: 15 October 2009 / Accepted: 26 February 2010 / Published online: 6 August 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Appropriate combinations of immunotherapy
and gene therapy promise to be more effective in the treat-
ment of cancer patients than either of these therapeutic
approaches alone. One such treatment is based on the appli-
cation of patients’ cytotoxic T cells, which can be activated,
expanded, and genetically engineered to recognize particular
tumor-associated antigens (TAAs). Because T cells recog-
nizing TAAs might become unresponsive in the process of
tumor development as a result of tumor evasion strategies,
immunogenic viral antigens or alloantigens could be used for
the expansion of cytotoxic T cells and then redirected through
genetic engineering. This therapeutic approach has already
demonstrated promising results in melanoma patients and
could be used in the treatment of many other tumors. The
graft-versus-leukemia, or more generally graft-versus-tumor,
reaction based on the application of a donor lymphocyte
infusion can also be ameliorated through the incorporation of
suicide genes into donor lymphocytes. Such lymphocytes
could be safely and more extensively used in tumor patients
because they could be eliminated should a severe graft-ver-
sus-host reaction develop.
Keywords Adoptive tumor immunotherapy
Gene therapy Graft-versus-tumor
Rebalancing Immune Specificity and Function in Cancer
by T-Cell Receptor Gene Therapy
Akshata Udyavar • Terrence L. Geiger
Received: 17 February 2010 / Accepted: 16 April 2010 / Published online: 1 August 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Adoptive immunotherapy with tumor-specific
T lymphocytes has demonstrated clinical benefit in some
cancers, particularly melanoma. Yet isolating and
expanding tumor-specific cells from patients is challenging
and there is limited ability to control T-cell affinity and
response characteristics. T-cell receptor (TCR) gene ther-
apy, in which T lymphocytes for immunotherapy are
redirected using an introduced rearranged TCR, has
emerged as an important alternative. Successful TCR gene
therapy requires consideration of a number of issues,
including TCR specificity and affinity, optimal gene ther-
apy constructs, types of T cells administered, and the
survival and activity of the modified cells. In this review
we highlight the rationale for and experience with TCR
gene therapy as well as new approaches to enhancing it.
Keywords T-cell receptor Adoptive immunotherapy
Cancer Gene therapy
Antimicrobial Peptides in the Brain
Yanhua Su • Kai Zhang • Hermann J. Schluesener
Received: 2 March 2010 / Accepted: 4 June 2010 / Published online: 29 July 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Antimicrobial peptides (AMPs) are an evolu-
tionarily conserved component of the innate immune
system of many species. The brain is an immunologically
privileged organ but can produce a robust immune
response against pathogens and cell debris, promoting
rapid and efficient clearance. AMPs may be critically
involved in the innate immune system of the brain. Though
the mechanisms of AMPs’ action in the brain still need
further elucidation, many studies have shown that AMPs
are multifunctional molecules in the brain. In addition to
antimicrobial action, they take part in congenital and
adaptive immune reactions (immunoregulation), function
as signaling molecules in tissue repair, inflammation and
other important processes through different mechanisms,
and they might, in addition, become diagnostic markers of
brain disease.
Keywords Antimicrobial peptides Brain
Antimicrobial activity Immunomodulatory activity
Signaling molecules
Biological Factors Involved in Implant-Anchored Orthodontics
and in Prosthetic-Implant Therapy: A Literature Review
Joanna Antoszewska • Katarzyna Raftowicz-Wo´jcik •
Beata Kawala • Teresa Matthews-Brzozowska
Received: 15 December 2009 / Accepted: 30 March 2010 / Published online: 30 July 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract During the past few years, the application of
orthodontic miniscrews and dental implants has been
expanded. However, failures have necessitated ongoing
investigation of potential risk factors. The aim of this
overview was to conduct an assessment of the immuno-
logical response following application of cortical temporary
anchorage devices—titanium miniscrew implants—in
orthodontic patients. A scrupulous search of the database
revealed only two matching items; therefore studies eval-
uating the immune response subsequent to insertion of
dental implants were reviewed. Thorough assessment
revealed the following as factors associated with dental
implant rejection: (1) correlation of the volume of gingival
and peri-implant crevicular fluid and the amount of inter-
leukin (IL)-1b with mucosal inflammation, thus serving as
a peri-implantitis evaluation index; (2) significantly more
frequent marginal bone loss around implants in patients
with IL-1B-511 2/2 genotype; (3) humoral response to
Staphylococcus aureus. However, since there is almost no
evidence-based evaluation of the allergic/inflammatory
reaction either to orthodontic titanium miniscrews them-
selves or in adolescents and young adults, who comprise
the largest group of orthodontic patients, this issue requires
further investigation. It is essential in order to achieve
successful, sophisticated and modern treatment of
malocclusions.
Keywords Orthodontics Miniscrew implants
Immunological response
The Incidence and Significance of Pattern-Recognition Receptors
in Chronic Viral Hepatitis Types B and C in Man
Iwona Mozer-Lisewska • Jan Sikora •
Arleta Kowala-Piaskowska • Mariusz Kaczmarek •
Grzegorz Dworacki • Jan _Zeromski
Received: 2 December 2009 / Accepted: 11 February 2010 / Published online: 10 June 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Chronic viral hepatitis B and C are among the
most common and devastating liver diseases worldwide.
Immune response plays a crucial role in the course of both
diseases. In spite of the importance of the adaptive arm of
the immune response, there is a growing role of innate
immunity, the earliest confronted with viral attack. Pattern-
recognition receptors (PRRs) and, in particular, Toll-like
receptors (TLRs) are molecules which are able not only to
recognize foreign invaders, but also quickly mount an
antiviral defense. Activation of PRRs has been demon-
strated in both hepatitis types, i.e. in situ in the liver and on
while blood cells. Both viruses, HCV and HBV, are able to
subvert the PRR-mediated antiviral response by means of
various proteins and enzymes. HCV acts via the non-
structural proteins NS2 and NS3/4A, while HBV HBeAg is
inversely correlated with TLR activity. Viral counterattack
is particularly directed toward dendritic cells, those creat-
ing the link with the adaptive immune response. Apart
from TLRs, other PRRs such as RIG-1 and MDA-5 are also
able to recognize viral infection and participate in the
activation of type I interferon synthesis. TLRs manifest
gene polymorphism, which was shown to affect several
consequences associated with chronic viral hepatitis such
as liver cirrhosis and the outcome of liver allotransplanta-
tion. There have been numerous attempts to take advantage
of the existence and activity of PRRs for the patients’
benefit. Several authors examined the role of TLR synthetic
agonists as inducers of TLR activation. In hepatitis C the
most promising agonists appear to be TLR3, 7, and 9 for
potential antiviral therapy. PRRs may also act as potent
adjuvants in HBV vaccines. Their baseline mRNA levels
may have predictive value in the course of antiviral
therapy.
Keywords Innate immunity
Pattern recognition receptors Viral hepatitis
Human Mast Cells and Mastocytosis: Harnessing MicroRNA
Expression as a New Approach to Therapy?
Lorenzo Deho’ • Silvia Monticelli
Received: 9 October 2009 / Accepted: 6 January 2010 / Published online: 29 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract MicroRNAs (miRNAs) are short, non-coding
RNAs that have emerged as key post-transcriptional regula-
tors in a wide variety of organisms and critical cellular
processes. Because any one miRNA can regulate the expres-
sion of a distinct set of genes, differential miRNA expression
can shape the repertoire of proteins that are actually expressed
during development, differentiation, or disease. To understand
what goes wrong when a cell becomes transformed requires
knowledge of the processes that ensure normal development.
It is now clear that miRNAs may act as oncogenes and/or
tumor suppressors within gene regulatory networks, thereby
contributing to the development of cancer. Mast cells are long-
lived cells, widely distributed throughout vascularized tissues,
in particular near surfaces that are exposed to the environment
(such as skin, airways, and the gastrointestinal tract), where
they contribute to bacterial clearance, enhancement of adap-
tive immune responses, modulation of inflammation, and the
degradation of toxic peptides and venoms. Here we review
current knowledge in the field of mast-cell differentiation and
disease in humans and mice and discuss future directions and
links between mast-cell differentiation, oncogenic transfor-
mation, and microRNAs as well as possible new points of
entry for therapeutic intervention.
Keywords Mast cells MicroRNA Mastocytosis
Gene regulation Disease
Intercellular Exchange of Surface Molecules and its Physiological
Relevance
Kathryn Brown • Mehmet Fidanboylu •
Wilson Wong
Received: 12 October 2009 / Accepted: 11 February 2010 / Published online: 28 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract For many decades, cellular immunologists have
relied on the expression of various cell surface molecules
to divide cells into different types and subtypes to study
their function. However, in recent years, a large and fast-
expanding body of work has described the transfer of
surface molecules, including MHC class I and II mole-
cules, between cells, both in vitro and in vivo. The function
of this process is still largely unknown, but it is likely to
have a significant role in the control of the immune system.
It is also likely that this process takes place in a regulated
rather than stochastic manner, thus providing another way
for the immune system to orchestrate its function. In this
review we will summarize the key findings so far, exam-
ining the mechanisms of transfer, the consequences of this
transfer as shown by in vitro experiments, and possible
consequences for the wider immune response.
Keywords Intercellular transfer MHC
Antigen presentation
Immunomodulatory Effects of IFN-b and Lovastatin
on Immunophenotype of Monocyte-Derived Dendritic
Cells in Multiple Sclerosis
Halina Bartosik-Psujek • Jacek Tabarkiewicz •
Krystyna Pocinska • Sebastian Radej •
Zbigniew Stelmasiak • Jacek Rolinski
Received: 1 September 2009 / Accepted: 1 March 2010 / Published online: 6 June 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Multiple sclerosis (MS) is an autoimmune
disease of the central nervous system and current MS
treatment is only partially effective. Recent data suggest
that statins may be potent immunomodulatory agents. In
order to evaluate their role in MS, we analyzed the in vitro
effects of interferon (IFN)-b and lovastatin on the differ-
entiation and maturation of monocyte-derived dendritic
cells (DCs) of MS patients. Twenty-seven patients with
relapsing–remitting MS were recruited for the study. DC
differentiation and maturation were evaluated based on
surface phenotypic changes and the expressions of CD14,
CD83, CD1a, CD80, CD86, CD206, and C209 were ana-
lyzed by flow cytometry. The results showed that IFN-b
and lovastatin affect DC phenotype. Both agents decrease
the expression of CD1a, which indicates a weakened pre-
sentation of glycolipid antigens. IFN-b causes up-regulated
and lovastatin down-regulated expression of CD86, which
results in a biased Th-cell responses in MS. Furthermore,
high doses of lovastatin cause a decrease in CD209
expression on the surface of DCs and can limit their
migration to various tissues. One of the mechanisms of the
beneficial action of IFN-b and statins may be associated
with their influence on DCs.
Keywords Dendritic cells Interferon beta Lovastatin
Multiple sclerosis
Hypoxia-inducible Factor 1 as one of the ‘‘Signaling Drivers’’
of Toll-like Receptor-Dependent and Allergic Inflammation
Vadim V. Sumbayev • Sally A. Nicholas
Received: 14 October 2009 / Accepted: 15 January 2010 / Published online: 26 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Hypoxia-inducible factor 1 (HIF-1) is a hete-
rodimeric transcription complex which plays a crucial role
in cellular adaptation to low oxygen availability. In the last
years there has been increasing evidence about the role of
this factor in inflammatory/innate immune reactions. It has
also been found to contribute to different types of allergic
inflammation. In this review the current knowledge about
the accumulation and role of HIF-1 in Toll-like receptor-
mediated and allergic inflammation is summarized. Dif-
ferential biochemical mechanisms employed to stabilize
the protein in different cases are discussed.
Keywords Inflammation Allergy Toll-like receptors
Hypoxia HIF-1
Recent Developments in Immune Activation by Uric Acid Crystals
Gilbert Ng • Elaine M. T. Chau • Yan Shi
Received: 20 August 2009 / Accepted: 11 February 2010 / Published online: 26 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Uric acid crystals, the causative agent of gout,
have recently gained widespread attention due to their role
as a natural endogenous adjuvant. Uric acid crystals, first
sensed extracellularly by membrane lipid alterations, are
internalized and subsequently activate the NLRP3 inflam-
masome. Currently, various aspects of this particular novel
pathway are poorly defined. This short review will focus on
some recent discoveries regarding this simple crystalline
structure and address areas requiring further investigation.
The fact that uric acid crystals activate innate host defense
mechanisms, triggering robust inflammation and immune
activation, may lead to engineering potent adjuvants for
future vaccines. Furthermore, the elucidation of uric acid’s
mechanism of inflammation may lay the foundation for
other solid inflammatory structures such as silica, asbestos,
and alum.
Keywords Uric acid Gout Adjuvant Vaccine
Lipid sorting NLRP3 inflammasome Inflammation
Impact of Human IgA Antibodies on Complement-Dependent
Cytotoxicity Mediated by Combinations of EGF-R-Directed
Antibodies
Stefan Lohse • Matthias Peipp • Thomas Beyer •
Thomas Valerius • Michael Dechant
Received: 9 October 2009 / Accepted: 11 January 2010 / Published online: 28 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Dual combinations of non-crossblocking
epidermal growth factor receptor (EGF-R)-directed
monoclonal antibodies were demonstrated to effectively
induce complement-dependent cytotoxicity (CDC) of
tumor cells, whereas individual antibodies were ineffec-
tive. Here the modulating effects of different antibody
isotypes on CDC were studied by adding them as a third
antibody. Two different combinations of non-crossblocking
EGF-R antibodies of human IgG1 isotype, 018/003 and
425/005, were investigated against the A431 and A1207
cell lines. As a third antibody, human IgG1, IgA1, and
IgA2 isotype variants of the therapeutic EGF-R antibody
225 were employed that bind to an EGF-R epitope distinct
from the other EGF-R antibodies. In this model, the human
IgG1 antibody proved to further enhance CDC, whereas
both IgA antibodies significantly blocked CDC. The IgG1
and IgA variants increased target opsonization at similar
levels, but the isotypes differed in their effects on C1q
fixation. Addition of IgG1 significantly enhanced comple-
ment factor binding on the target surface, whereas both IgA
antibodies reduced complement binding. Control experi-
ments revealed this blocking effect to be not specific to IgA
antibodies, but to antibody constructs incapable of acti-
vating the complement system. Interestingly, the effects
caused by the IgA2 isotype were consistently stronger than
those by IgA1, which may be caused by stronger steric
hindrance due to its reduced hinge flexibility. These results
demonstrate that monoclonal IgA antibodies inhibit IgG-
mediated complement activation in vitro and suggest that
the appearance of IgA antibodies within a polyclonal
immune response might inhibit complement activation
in vivo.
Keywords IgA Complement CDC
Don’t Get Spooked! How to Collaborate with a Professional
Medical Communicator (And Avoid Ghostwriting)
Cindy W. Hamilton
Received: 22 April 2010 / Accepted: 4 May 2010 / Published online: 28 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Substantial confusion exists about the role of
medical writers and editors (hereafter, medical communi-
cators) in medical publication. Much of the confusion is due
to the failure to recognize the difference between terms.
Ghostwriting is unethical, whereas professional medical
communication refers to legitimate writing and editing ser-
vices provided by individuals who comply with ethical
guidelines. The purpose of this article is to shed light on this
subject by reviewing relevant guidelines and by providing
practical tips for authors interested in collaborating with
medical communicators. Specifically, this article addresses a
series of questions, such as what to expect from medical
communicators, how to evaluate them, and how to collabo-
rate ethically and efficiently with them. To ensure that the
process is ethical, authors should begin collaborating with
the medical communicator early in the process, continue
doing so throughout manuscript development, and control
manuscript content. In addition, authors should disclose
substantial contributions and funding sources of the medical
communicator and all other individuals not meeting
authorship criteria. To ensure that the process is efficient,
authors should delegate time-consuming technical writing
and editing tasks to the medical communicator.
Keywords Interprofessional relations
Medical education Professional ethics Writing standards
Slime Production by Staphylococcus aureus and Staphylococcus
epidermidis Strains Isolated from Patients with Diabetic Foot
Ulcers
Adrianna Podbielska • Hanna Galkowska •
Ewa Stelmach • Grazyna Mlynarczyk •
Waldemar L. Olszewski
Received: 24 September 2009 / Accepted: 9 December 2009 / Published online: 26 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Slime production is a very important factor
related to biofilm formation. The objective of the present
study was to determine the frequency of slime production
by Staphylococcus aureus and Staphylococcus epidermidis
strains recovered from 50 patients with diabetic foot ulcers.
Slime production was determined using the Congo red agar
(CRA) method and compared with immunocytochemistry
for the production of polysaccharide intercellular adhesin
(PIA). Out of 55 S. aureus strains, 69% produced slime as
shown by the CRA method. Of them, 84.2% also produced
PIA. Of 17 CRA-negative strains, 70.6% produced PIA.
Out of 20 S. epidermidis strains, 75% were CRA positive
and 93.3% produced PIA. All CRA-negative S. epidermidis
produced PIA. In conclusion, PIA production is a very
common trait of S. aureus and S. epidermidis isolates
obtained from diabetic foot ulcer patients.
Keywords Slime production Staphylococci
Polysaccharide intercellular adhesin
Erratum to: Complement: Coming Full Circle
Gae¨lle Le Friec • Claudia Kemper
Published online: 10 April 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Erratum to: Arch Immunol Ther Exp (2009) 57:393–407
DOI 10.1007/s00005-009-0047-4
The elements of the first author’s name were wrongly
coded in the metadata of this article, with the result that the
name was listed incorrectly in all indexing services. Her
given name is Gae¨lle; her surname, Le Friec.
Experimental Anticancer Therapy with Vascular-disruptive
Peptide and Liposome-entrapped Chemotherapeutic Agent
Aleksander Sochanik • Iwona Mitrus •
Ryszard Smolarczyk • Tomasz Cichon´ •
Mirosław S´nietura • Maria Czaja • Stanisław Szala
Received: 22 May 2009 / Accepted: 29 October 2009 / Published online: 11 April 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Vasculature is essential for the sustained growth
of solid tumors and metastases. Tumor cells surviving vas-
cular-disruptive therapeutic intervention (especially those
present at the tumor rim) can contribute to tumor regrowth.
The aim was to strengthen, by carrier-mediated delivery of a
chemotherapeutic, the curative effects of a bifunctional anti-
vascular oligopeptide capable of inducing vascular shut-
down and tumor shrinkage. For the in vitro experiments
and animal therapy, ACDCRGDCFC-GG-D(KLAKLAK)2
peptide (900 lM in D-PBSA, i.e. Dulbecco’s PBS without
Ca2? and Mg2?) and size-calibrated, passively or actively
targeted liposomes based on distearoylphosphatidylcholine,
cholesterol, and N-carbamoyl-methoxypolyethyleneglycol
coupled to distearoylphosphatidylethanolamine (PEG–
DSPE) and containing gradient-entrapped doxorubicin were
used. The KB (human nasopharyngeal carcinoma) cell line
overexpressing folate receptors was used in the fluorescence
studies of liposomal uptake. The B16-F10 melanoma cell
line was used for confirming, by flow cytometry and
confocal microscopy, doxorubicin intracellular transfer as
well as to induce experimental tumors in C57BL/6 mice.
Animal therapy was achieved with injections of vascular-
disrupting peptide, doxorubicin-loaded liposomes, or alter-
nating combined therapy. The results (tumor growth
inhibition and survival) were compared using the Mann–
Whitney U test and the log-rank test. Necrosis in H&E-
stained tumor sections was assessed microscopically by
pathologists. Treatment of C57BL/6 mice bearing B16-F10
experimental tumors with a combination of vascular-
disruptive peptide and doxorubicin-carrying pegylated
liposomes (either passively targeted liposomes (PTL) or
folate receptor targeted) gave better therapeutic effects when
tumor development was re-challenged with a second cycle
of combined therapy. Marked inhibition of tumor growth
and a statistically significant extension of the lifespan of the
treated mice were observed when the re-challenge involved
the use of folate receptor-targeted liposomes (FTL). Anti-
cancer therapy involving vascular-disruptive peptide and
doxorubicin delivered via pegylated folate receptor-targeted
liposomes is more effective than either monotherapy,
especially when tumor growth is re-challenged with the
therapeutic combination.
Keywords Vascular-disruptive agents
Pegylated liposomes Doxorubicin
Combined antitumor therapy
Comparison of Three Methods of DNA Extraction
in Endocervical Specimens for Chlamydia trachomatis Infection
by Spectrophotometry, Agarose Gel, and PCR
Anahita Jenab • Rasoul Roghanian •
Naser Golbang • Pouran Golbang •
Leili Chamani-Tabriz
Received: 9 April 2009 / Accepted: 4 February 2010 / Published online: 17 April 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Chlamydia trachomatis is the major cause of
sexually transmitted disease in the world. The aim of this
study was to determine the best method of DNA extraction
for detecting C. trachomatis by polymerase chain reaction
(PCR) in sexually active women (n = 80) attending Shahid
Beheshti Hospital in Isfahan, Iran. Endocervical swabs
were collected from 80 women, 22 of whom were
asymptomatic and 58 symptomatic. Three different DNA
extraction methods were used in this study (phenol-
chlorophorm, proteinase K, and boiling). DNA yield was
evaluated by spectrophotometry, agarose gel, and PCR.
The internal control was assayed by b-globin primers
(PCO4, GH20). The DNA cryptic plasmid was selected as
the target for C. trachomatis and samples were examined
by PCR using specific KL1 and KL2 primers. It was shown
that DNA extraction by boiling was the most sensitive with
the highest yield of DNA. Of the 80 samples, 17 (21.25%)
showed positivity for C. trachomatis by PCR. The highest
rate of C. trachomatis infection was found in the group
aged between 35 and 45 years old and those who used
withdrawal or an intrauterine device as methods of con-
traception. It was demonstrated that DNA extraction by
boiling was the least expensive and a very rapid method
that gave the highest DNA yield. The infection rate in the
sexually active women, including symptomatic and
asymptomatic, was 21.25%, with a presumably high
prevalence compared with other studies done in this field.
Keywords Chlamydia trachomatis
Endocervical smears DNA extraction PCR
Agarose gel Spectrophotometry
Regulatory T Cell as a Target for Cancer Therapy
Lucas Cunha Dias de Rezende •
Ian Victor Silva • Letı´cia Batista Azevedo Rangel •
Marco Cesar Cunegundes Guimara˜es
Received: 23 September 2009 / Accepted: 6 December 2009 / Published online: 7 April 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Advances in our understanding of CD4?
CD25?Foxp3? regulatory T cells (TRegs) enabled the
characterization of their activities in maintaining peripheral
tolerance, preventing autoimmune diseases, and limiting
chronic inflammatory diseases. Ironically, an effective
action of these cells during tumor development can limit
beneficial responses by suppressing immunity and limiting
antitumor resistance, whereas one of the main functions of
the immune system is to eliminate malignant cells. During
the last years, the immunological role, mechanism of
action, and clinical importance of these cells were pro-
foundly characterized and the relationship between this
subset of lymphocytes and cancerous cells arises as a key
factor that influences tumor development. Recent insights
obtained from clinical studies and experimental mouse
models expand our perception of the potential role of TRegs
in cancer treatment. In this review we describe the basic
mechanisms of TReg origin and differentiation, their
potential role in cancer, as well as the future perspectives
concerning the modulation of these cells as a potential
approach for anticancer strategies.
Keywords Cancer Immunotherapy
Regulatory lymphocytes Foxp3
Adoptive T-cell Immunotherapy of Cancer Using Chimeric
Antigen Receptor-Grafted T Cells
David Marc Davies • John Maher
Received: 1 June 2009 / Accepted: 27 October 2009 / Published online: 6 April 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Harnessing the power of the immune system to
target cancer has long been a goal of tumor immunologists.
One avenue under investigation is the modification of
T cells to express a chimeric antigen receptor (CAR).
Expression of such a receptor enables T-cell specificity to
be redirected against a chosen tumor antigen. Substantial
research in this field has been carried out, incorporating a
wide variety of malignancies and tumor-associated anti-
gens. Ongoing investigations will ensure this area continues
to expand at a rapid pace. This review will explain the
evolution of CAR technology over the last two decades in
addition to detailing the associated benefits and disadvan-
tages. The outcome of recent phase I clinical trials and the
impact that these have had upon the direction of future
research in this field will also be addressed.
Keywords Chimeric antigen receptor T-cell
Cancer Adoptive transfer Survival
B Cells: From Early Development to Regulating Allergic Diseases
Konstantinos Samitas • Jan Lo¨tvall •
Apostolos Bossios
Received: 3 July 2009 / Accepted: 27 October 2009 / Published online: 11 May 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract B lymphocytes are characterized by a unique
and highly specialized developmental pathway that is
responsible for their vast phenotypic and function diversity.
B cell development is strictly regulated to ensure sufficient
specific humoral immunity while at the same time avoiding
any errors that would compromise B cell functionality. The
generation and maintenance of mature B cells from the
constant flux of bone marrow progenitors is a complex
process that is generally poorly understood, although great
progress has been made in recent years. B cells have for
long been considered mainly as antibody-producing cells
and therefore believed to play an important role in the
pathophysiology of allergic diseases, primarily through
their ability to produce IgE antibodies. However, recent
findings have revealed new aspects of their role in immune
responses that place them again under the spotlight as
important immune regulators, independent of antibody
production. This review focuses on the developmental
processes responsible for the numerous phenotypes and
functions of the B-lymphocyte pool and the different
aspects of effector B cell functionality in the context of
allergy.
Keywords B cell development Bone marrow
Regulation Allergy Effector B cell
Involvement of Envelope-Glycoprotein Glycans
in HIV-1 Biology and Infection
Milan Raska • Jan Novak
Received: 24 September 2009 / Accepted: 30 November 2009 / Published online: 6 April 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Infection of host cells with HIV-1 depends on a
highly glycosylated virus envelope glycoprotein (Env) and
host-cell receptors. Glycans participate substantially in Env
folding and in the binding of virions to the host-cell surface
and indirectly affect cellular uptake of HIV-1. Moreover,
Env glycans could protect HIV-1 from host’s neutralizing
antibodies, but some glycans, on the other hand, represent
targets for neutralizing antibodies. Variability of Env and its
glycans in the HIV-1 strains from around the world as well
as in patients during disease progression contributes sub-
stantially to further HIV-1 spreading in spite of the progress
in basic HIV-1 research, vaccine development, and highly
active antiretroviral therapy of HIV-1 infections.
Keywords HIV-1 gp120 glycosylation
gp120 folding gp120 antibody recognition
Microbiological Findings and Treatment of EBV-Associated
Hemophagocytic Lymphohistiocytosis: A Case Report
Maciej Przybylski • Tomasz Dziecia˛tkowski •
Dorota Zdun´ czyk • Wiesław Wiktor Je˛drzejczak •
Mirosław Łuczak
Received: 30 June 2009 / Accepted: 11 August 2009 / Published online: 13 April 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Epstein–Barr virus (EBV) is the major triggering
factor for hemophagocytic syndrome or hemophagocytic
lymphohistiocytosis (HLH). In patients with EBV-HLH, the
EBV-infected T cells or natural killer cells are mostly mono-
or oligoclonally proliferating, whereby hypercytokinemia
plays a major role and causes hemophagocytosis, cellular
damage, and dysfunction of various organs. This report
describes the detection and treatment of EBV-associated
HLH in the case of a 17-year-old male. Serum samples and
skin swabs were tested for the presence of viral DNA using
real-time PCR techniques. To confirm the molecular bio-
logical tests, electron microscopy was also performed. EBV
DNA was detected with real-time PCR in both blood sam-
ples and skin swabs. The level of viral DNA constantly
decreased during the applied therapy. The presence of the
virus in the skin was confirmed by the appearance of herpes
virus-like particles detected by electron microscopy in fluid
taken from skin ulcerations. The results show that in terms of
treatment, special therapeutic measures are required to
control the cytokine storm generated by EBV and to suppress
proliferating EBV genome-containing cells because the
clinical course is often fulminate and results in a poor
outcome. Therefore the potential of chemotherapy with
a combination of steroids, etoposide, and cyclosporine to
control HLH was assessed in the adolescent, who met the
stringent diagnostic criteria for this reactive disorder of the
mononuclear phagocyte system.
Keywords Epstein–Barr virus Hemophagocytic
lymphohistiocytosis Real-time PCR
Dysregulation of the Receptor Activator of NF-jB Ligand
and Osteoprotegerin Production Influence the Apoptosis
of Multiple Myeloma Patients’ Bone Marrow Stromal Cells
Co-Cultured with Myeloma Cells
Barbara Zdzisin´ ska • Agnieszka Bojarska-Junak •
Adam Walter-Croneck • Martyna Kandefer-Szerszen´
Received: 25 February 2009 / Accepted: 2 July 2009 / Published online: 16 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract The interaction of multiple myeloma (MM)
cells and bone marrow stromal cells (BMSCs) induces
profound changes in the bone marrow environment,
influencing osteoclastogenesis and MM cell survival. Dif-
ferences in receptor activator of NF-jB ligand (RANKL)
and osteoprotegerin (OPG) production in BMSCs derived
from MM patients and control subjects and the apoptosis of
BMSCs and MM cells in co-cultures of both cell types were
examined. RANKL and OPG expressions were examined
by ELISA and semiquantitative RT-PCR. Apoptosis of
BMSCs after contact with RPMI8226 and U266 cells was
measured by flow cytometry and the level of ALP activity
by the spectrophotometric method. OPG production by
BMSCs was significantly inhibited after direct contact with
RPMI8226 cells. Production of soluble RANKL was
enhanced and the increase was more significant in the
BMSCs of the MM patients than in those of the controls. In
co-cultures of BMSCs and MM cells, significant apoptosis
was detected with a concomitant decrease in ALP activity.
This apoptosis decreased significantly in the presence of
RANK-Fc, an antagonist of RANKL. Disturbances in the
RANKL/OPG system are more profound in the BMSCs of
MM patients than in those of control subjects after direct
contact with RPMI8226 cells. Moreover, direct contact with
RPMI8226 and U266 cells induces apoptosis of BMSCs
which is mediated by an overproduction of RANKL.
Keywords Multiple myeloma
Bone marrow stromal cells RANKL OPG Apoptosis
P2X7 Receptor Signaling Pathway as a Therapeutic Target
for Neurodegenerative Diseases
Takato Takenouchi • Kazunari Sekiyama • Akio Sekigawa •
Masayo Fujita • Masaaki Waragai • Shuei Sugama •
Yoshifumi Iwamaru • Hiroshi Kitani • Makoto Hashimoto
Received: 2 September 2009 / Accepted: 12 November 2009 / Published online: 9 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract A recent study suggested that neuroinflamma-
tion plays a major role in the pathogenesis of a number of
neurodegenerative diseases such as Alzheimer’s disease
and Parkinson’s disease. Although the precise mechanism
is obscure, dysregulation of the signaling transduction
pathway in microglia may enhance inflammation, leading
to synaptic dysfunction and ultimately to neuronal cell
death. The expression and function of the P2X7 receptor
(P2X7R), an ATP-gated ion channel abundantly expressed
in microglia in the brain, is significantly up-regulated in the
postmortem brain of Alzheimer’s disease patients and
various neurodegenerative disease animal models. This
supports the role of the P2X7R pathway in the progression
of neurodegeneration. Blocking P2X7R using brilliant blue
G, a P2X7R antagonist that can cross the blood–brain
barrier, has been shown to result in the amelioration of
neuropathology in various animal models. Taken together,
these results raise the possibility that the P2X7R signaling
pathway could be a therapeutic target for treating various
neurodegenerative diseases.
Keywords P2X7 receptor Microglia
Neuroinflammation Neurodegenerative disease
Immunoprophylaxis of Tuberculosis:
An Update of Emerging Trends
Neelja Singhal • Deepa Bisht • Beenu Joshi
Received: 14 April 2009 / Accepted: 6 July 2009 / Published online: 7 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Developing effective prophylactics to combat
tuberculosis is currently in an exploratory stage. The HIV
pandemic and emergence of multi- and extensively drug-
resistant strains of Mycobacterium tuberculosis indicate
that the current preventive measures against this ever-
evolving pathogen are inadequate. The currently available
vaccine BCG in its present form affords variable protection
which usually wanes with aging. Various reasons have
been cited to explain the discrepancies in the efficacy of
BCG, including generic differences in the different BCG
vaccine strains used in immunization program throughout
the world. The low efficacy of BCG vaccine has promoted
the search for novel vaccines for tuberculosis. The search
strategies aim at completely replacing the existing vaccine
and/or augmenting/improving the current BCG vaccine.
Among new vaccine candidates are live attenuated
M. tuberculosis vaccines, recombinant BCG, DNA vaccines,
subunit vaccine, and fusion protein-based vaccines. More
than 200 new vaccine candidates have been developed as a
result of research work over the past few years. To date, at
least eight vaccine candidates are undergoing clinical
evaluation, with a few of them successfully qualifying in
the first phase of clinical testing. These recent advances
present an optimistic insight whereby a new tuberculosis
vaccine might be expected to be available for public use in
the next few years.
Keywords BCG M. tuberculosis Vaccine
Prophylactics
CD4+ T Cell Epitope Discovery and Rational Vaccine Design
Daniela Santoro Rosa • Susan Pereira Ribeiro •
Edecio Cunha-Neto
Received: 16 June 2009 / Accepted: 8 August 2009 / Published online: 14 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract T cell epitope-driven vaccine design employs
bioinformatic algorithms to identify potential targets of
vaccines against infectious diseases or cancer. Potential
epitopes can be identified with major histocompatibility
complex (MHC)-binding algorithms, and the ability to bind
to MHC class I or class II indicates a predominantly CD4?
or CD8? T cell response. Furthermore, an epitope-based
vaccine can circumvent evolutionary events favoring
immune escape present in native proteins from pathogens.
It can also focus on only the most relevant epitopes (i.e.
conserved and promiscuous) recognized by the majority of
the target population. Mounting evidence points to the
critical role of CD4? T cells in natural antigen encounter
and active immunization. In this paper the need for CD4?
T cell help in vaccine development, the selection of CD4?
T cell epitopes for an epitope-based vaccine, and how the
approach can be used to induce a protective effect are
reviewed.
Keywords Vaccine design CD4? T cells T cell help
Epitope prediction Epitope-based vaccines
Plasma TNF-a and IL-10 Level-Based Prognostic Model Predicts
Outcome of Patients with Diffuse Large B-Cell Lymphoma
in Different Risk Groups Defined by the International
Prognostic Index
Ewa Lech-Maranda • Jacques Bienvenu • Florence Broussais-Guillaumot •
Krzysztof Warzocha • Anne-Sophie Michallet • Tadeusz Robak •
Bertrand Coiffier • Gilles Salles
Received: 17 November 2008 / Accepted: 20 July 2009 / Published online: 27 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Tumor necrosis factor (TNF)-a and interleukin
(IL)-10 are key cytokines involved in lymphoma devel-
opment. Their pretreatment plasma levels were reported to
influence the clinical course of non-Hodgkin’s lymphoma.
In this study the impact of combined elevation of TNF-a
and IL-10 on disease features and outcome of patients with
diffuse large B-cell lymphoma (DLBCL) were investi-
gated. Plasma TNF-a and IL-10 levels were determined at
the time of diagnosis in a group of 106 DLBCL patients
uniformly treated with anthracycline-based regimens.
Three risk groups depending on the pretreatment levels of
the cytokines were identified: low-, intermediate-, and
high-risk groups. In univariate analysis, the cytokine
intermediate- and high-risk groups were associated with
lower probability of achieving a complete remission (odds
ratio [OR] = 0.2, 95% confidence interval [CI] 0.06–0.6,
p = 0.006 and OR = 0.05, 95% CI 0.01–0.2, p \ 0.0001,
respectively) and shorter progression-free survival (PFS)
(OR = 4.4, 95% CI 1.9–10.2, p \ 0.001 and OR = 9.7,
95% CI 4.1–23.0, p \ 0.0001, respectively) and overall
survival (OS) (OR = 4.2, 95% CI 1.7–10.1, p = 0.002 and
OR = 11.2, 95% CI 4.4–28.4, p \ 0.0001, respectively) in
comparison with the cytokine low-risk group. In multi-
variate analysis, the cytokine intermediate- and high-risk
groups also correlated with shorter PFS (relative risk
[RR] = 4.5, 95% CI 1.9–10.9, p = 0.001 and RR = 5.8,
95% CI 2.2–15.3, p \ 0.0001, respectively) and OS
(RR = 4.6, 95% CI 1.8–12.0, p = 0.001 and RR = 7.5,
95% CI 2.7–20.9, p \ 0.0001, respectively) regardless of
the International Prognostic Index (IPI) scoring system.
The TNF-a and IL-10 level-based index may work as an
additional model to the IPI for predicting the survival of
DLBCL patients. This model may help to identify patients
in a given IPI risk group for whom more accurate and risk-
adapted treatment could be advised.
Keywords Tumor necrosis factor a Interleukin-10
International Prognostic Index
Diffuse large B-cell lymphoma
Nanogram Doses of Alum-Adjuvanted HBs Antigen Induce
Humoral Immune Response in Mice When Orally Administered
Jo´zef Kapusta • Tomasz Pniewski •
Jacek Wojciechowicz • Piotr Bocia˛g •
Andrzej Płucienniczak
Received: 25 February 2009 / Accepted: 4 August 2009 / Published online: 18 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Mucosal immunity elicited by plant-based and
other orally administered vaccines can serve as the first line
of defense against most pathogens infecting through
mucosal surfaces, but it is also considered for systemic
immunity against blood–borne diseases such as hepatitis B
(HB). Previous oral immunization trials based on multiple
administration of high doses of HBs antigen elicited an
immune response; however, a reproducible and long-lasting
immunization protocol was difficult to design. The objec-
tive of this study was to evaluate the effect of dose and
timing of orally delivered alum-adsorbed antigen on the
magnitude of the anti-HBs humoral response. Mice were
immunized orally by gavage intubation or parenterally by
intramuscular injection three times, once every 2 weeks,
with doses of 5, 50, or 500 ng alum-adjuvanted HBsAg. A
low dose (10 ng) of HBsAg was orally administered three
times in different time intervals: 2, 4, 6, and 8 weeks. The
three consecutive 5-ng oral doses of the antigen induced
immune response at the protective level (C10 mIU/ml),
significantly higher than the reaction elicited by three 50 or
500 ng doses. In contrast, intramuscular delivery of these
doses did not differ significantly; however, they induced a
five to six times higher immune response than oral immu-
nization. The 8-week period between each of the three oral
immunizations appeared to be favorable to the anti-HBs
humoral responses compared with the shorter schedules.
The results presented here clearly identify the importance of
low doses of antigen administered orally in extended
intervals for a significantly higher anti-HBs response. This
finding provides some indications concerning the strategy
of orally administered vaccines, including plant-based ones.
Keywords HBs antigen HBsAg Oral immunization
Anti-HBV oral vaccine
The Role of Thymic Stromal Lymphopoietin in Allergic
Inflammation and Chronic Obstructive Pulmonary Disease
Cailong Fang • Leonard Q. C. Siew •
Chris J. Corrigan • Sun Ying
Received: 15 May 2009 / Accepted: 6 August 2009 / Published online: 9 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract Thymic stromal lymphopoietin (TSLP) primes
dendritic cells to promote a Th2 inflammatory response. Its
action is mediated by a heterodimeric receptor which
consists of the interleukin-7 receptor a chain and the TSLP
receptor chain (TSLPR). TSLPR resembles the common c
chain subunit utilized by many type 1 cytokine receptors.
Normal epithelial cells, keratinocytes, and stromal cells
constitutively express TSLP. Dendritic cells that are acti-
vated by TSLP promote the development of CD4? T cells
into pro-inflammatory Th2 cells. TSLP thus plays a
potentially important role in the pathogenesis of allergic
inflammation in asthma and atopic dermatitis. TSLP also
has direct effects on other types of cells in the bronchial
mucosa. It is over-expressed in the bronchial mucosa in
chronic obstructive pulmonary disease (COPD), which is
traditionally described as a Th1-related disease, as well as
severe asthma, which is traditionally described as a Th2-
related disease. In this review we will discuss TSLP
expression, function, and available and potential mecha-
nisms in both allergic inflammation and COPD.
Keywords TSLP Asthma COPD
Lentiviral Vectors in Gene Therapy: Their Current Status
and Future Potential
David Escors • Karine Breckpot
Received: 14 July 2009 / Accepted: 6 October 2009 / Published online: 9 February 2010
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2010
Abstract The concept of gene therapy originated in the
mid twentieth century and was perceived as a revolutionary
technology with the promise to cure almost any disease of
which the molecular basis was understood. Since then,
several gene vectors have been developed and the feasi-
bility of gene therapy has been shown in many animal
models of human disease. However, clinical efficacy could
not be demonstrated until the beginning of the new century
in a small-scale clinical trial curing an otherwise fatal
immunodeficiency disorder in children. This first success,
achieved after retroviral therapy, was later overshadowed
by the occurrence of vector-related leukemia in a signifi-
cant number of the treated children, demonstrating that the
future success of gene therapy depends on our under-
standing of vector biology. This has led to the development
of later-generation vectors with improved efficiency,
specificity, and safety. Amongst these are HIV-1 lentivirus-
based vectors (lentivectors), which are being increasingly
used in basic and applied research. Human gene therapy
clinical trials are currently underway using lentivectors in a
wide range of human diseases. The intention of this review
is to describe the main scientific steps leading to the
engineering of HIV-1 lentiviral vectors and place them in
the context of current human gene therapy.
Keywords Gene therapy Lentivirus-based vector