bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular heterogeneity of auto‐anti‐idiotypic antibodies in MLR‐lpr/lpr mice
Authors: Koizumi Tamio , Puccetti Antonio , Migliorini Paola , Barrett Kathleen J. , Schwartz Robert S.
Published: March 2007

The VH and Vx gene families expressed by 20 monoclonal auto‐anti‐idiotypes (Ab2) derived from unmanipulated MLR‐lpr/lpr mice were determined by Northern blotting. Complete variable region sequences of six Ab2, along with three additional Vx‐JH Ab2 sequences, were obtained. These auto‐anti‐idiotypes arose spontaneously in the animals, and they bound specifically to an idiotypic determinant (ld/r) on mAb 28/12, a monoclonal IgG2b MLR‐lpr/lpr anti‐small nuclear ribonucleoprotein antibody. The 16 Ab2 heavy chains belonged to 7 different VH gene families, and the 10 Ab2 light chains were derived from 8 Vx families. The light chains of two Ab2 were ≈99% identical; the remaining variable region sequences were highly heterogeneous. There was no correlation between primary amino acid sequence of either heavy or light chain and idiotypic properties of the auto‐anti‐idiotypes. Six Ab2 used VH or Vx genes that are identical to known germ‐line genes. A high proportion of the spontaneous auto‐anti‐idiotypes was shown to have autoantibody activity (anti‐DNA, anti‐ribonucleoprotein), or specific binding reactions with lipopolysaccharide of Salmonella RE, or both properties. The structural diversity of spontaneous MLR‐lpr/lpr auto‐anti‐idiotypes differs sharply from the structural homogeneity reported for Ab2 induced in normal animals against syngeneic Ab1. Our results suggest that auto‐anti‐idiotypes might arise independently of an immunogenic stimulus from an Ab1.
The T cell receptor/CD3 complex and CD2 stimulate the tyrosine phosphorylation of indistinguishable patterns of polypeptides in the human T leukemic cell line Jurkat
Authors: Ley Steven C. , Davies Adelina A. , Druker Brian , Crumpton Michael J.
Published: March 2007

Stimulation of the T cell receptor (TcR)/CD3 complex of Jurkat T cells with a monoclonal antibody to the CD3 ϵ chain induced the tyrosine phosphorylation of multiple polypeptides, ranging in size from 21 to 155 kDa. The protein tyrosine phosphorylation was characterized by its rapidity and its transient nature, returning to baseline levels by 60 min. Protein tyrosine kinase activity was also induced when the Jurkat T cells were stimulated with a mitogenic pair of antibodies directed against CD2. Comparison of the polypeptides which were phosphorylated on tyrosine in response to stimulation of the two receptors, by either one‐ or two‐dimensional analysis, failed to reveal any differences. These data suggest that the TcR/CD3 complex and CD2 activated the same tyrosine kinase or kinases. A model is proposed in which CD2 functions as a signal amplifier in physiological responses to antigen/major histocompatibility complex without changing the qualitative nature of the signal generated via the TcR/CD3 complex.
Bone marrow‐derived progenitor T cells convey the origin of maturational arrest from CD4+CD8+ to CD4−CD8+ thymocytes in LEC mutant rats
Authors: Agui Takashi , Sakai Tohru , Himeno Kunisuke , Matsumoto Kozo
Published: March 2007

A mutant strain of rats, LEC, shows a novel arrest of T cell maturation from CD4+CD8+ to CD4+CD8− but not to CD4−CD8+ cells in the thymus. Transplantation of LEC rat fetal thymuses into the subcapsule of the kidney of athymic nude rats resulted in a normal maturation of thymocytes in the thymus graft. Furthermore, both single‐positive thymocytes and peripheral lymph node T cells expressed T cell receptor α/β antigen, and lymph node T cells acquired the ability to produce interleukin 2 upon mitogen stimulation. Transplantation of fetal thymuses from LEA rats, which express the same major histocompatibility complex haplotype as LEC rats, into LEC rat kidney subcapsule resulted in the maturational arrest from CD4+CD8+ to CD4+CD8− cells in the thymus graft. These data strongly suggest that bone marrow‐derived progenitor T cells carry the cause of maturational arrest and that the thymic stroma of LEC rats has a normal potential to nurse thymocytes.
A B cell population that dominates the primary response to influenza virus hemagglutinin does not participate in the memory response
Authors: Kavaler Joshua , Caton Andrew J. , Staudt Louis M. , Gerhard Walter
Published: March 2007

The early primary B cell response of BALB/c mice to the influenza virus A/PR/8/34 (PR8) hemagglutinin (HA) is dominated by B cells that utilize a single Vx gene in association with one of two closely related VH genes. We have used an anti‐idiotypic reagent that recognizes a light chain‐associated idiotope (23‐1 Id) on these antibodies to follow their presence during the anti‐HA response. Quantitation of 23‐1 Id+ antibodies at different time points during the anti‐HA response indicates that the 23‐1 Id+ B cell response peaks early after primary immunization and is not re‐induced by secondary challenge. Furthermore, 23‐1 Id+ titers in serum decay rapidly between the first and second week after immunization, and the HA‐specific 23‐1 Id+ precursor B cell population does not significantly expand in the months following immunization. These results indicate that despite their predominance during the primary response, 23‐Id+ B cells abruptly disappear from the response and do not mature into memory B cells.
Biased T cell receptor Vα region repertoire in the synovial fluid of rheumatoid arthritis patients
Authors: Pluschke Gerd , Ricken Gesa , Taube Heiko , Kroninger Susanne , Melchers Inga , Peter Hans Hartmut , Eichmann Klaus , Krawinkel Ulrich
Published: March 2007

Synovial T lymphocytes seem to contribute to the pathogenesis of rheumatoid arthritis (RA). Since very little is known about the structural heterogeneity of their T cell antigen receptors (TcR), we analyzed TcR α chain mRNA of synovial fluid T cells from two RA patients. TcR α chain cDNA was amplified by the polymerase chain reaction with single‐sided specificity for the α chain constant (Cα) gene segment, and the nucleotide sequences of 51 functionally rearranged cDNA clones were determined. Twenty different Vα genes and 26 different Jα gene segments were utilized in these cDNA clones. Three of the Vα gene segments which are frequently (8%‐17% total) expressed in synovial fluid T cells have rarely been found in the TcR repertoire of peripheral blood T cells from healthy individuals. The T cell responses in the rheumatic synovia analyzed here are not oligoclonal, but the usage of TcR Vα genes is biased.
Induction of CD8 molecules on thymic γ/δ T cells in vitro is dependent upon α/β T cells
Authors: Spetz Anna‐Lena , Kourilsky Philippe , Larsson‐Sciard Eva‐Lotta
Published: March 2007

Thymocytes differentiate upon interactions with microenvironmental components, but the precise role of different stromal cells, or other T cells, in early differentiative events remains unclear. Here we have analyzed the in vitro differentiation of double‐negative (DN) thymocytes from young adult mice. We demonstrate that a substantial proportion of DN thymocytes differentiate into CD8+ γ/δ T cells upon stimulation with concanavalin A and recombinant interleukin 2. However, if α/β T cells are excluded from the initial population of DN thymocytes, the CD8+ γ/δ T cells do not appear in the cultures. These results suggest a role for T‐T cell interactions in thymic differentiative events, and provide evidence for physiological interactions between the α/β and γ/δ T cell compartments within the thymus.
Temperature‐induced stress abrogates co‐stimulatory function in antigen‐presenting cells
Authors: Kuperberg Gina , Ellis Joanna , Marcinkiewicz Janosz , Chain Benjamin M.
Published: March 2007

This study examines the effect of heat‐induced stress on the function of splenic antigen‐presenting cells. Heat shock (at 41°C, 45°C) profoundly inhibits the ability of this population to stimulate antigen‐specific proliferation in ex vivo, or short‐term cultures of T cells. This inhibition is not due to the release of suppressor factors from the antigen‐presenting cells, nor to a lack of interleukin 1. Comparison of the effect of heat shock on presentation to a T cell hybridoma suggest that heat shock may affect the ability of antigen‐presenting cells to deliver a co‐stimulatory signal to the T cell. The implication of these findings in terms of the regulation of autoimmune phenomena is discussed.
Antigen presentation by dendritic cells provides optimal stimulation for the production of interleukin (IL) 2, IL 4 and interferon‐γ by allogeneic T cells
Authors: Ellis Joanna , Chain Benjamin M. , Hugh Davies D. , Ibrahim Mohammed A. A. , Katz David R. , Kaye Paul M. , Lightstone Elizabeth
Published: March 2007

Previous studies have shown that dendritic cells are the most potent inducers of T cell proliferation in vitro and that this is reflected in the release of interleukin (IL) 2 into culture supernatants during dendritic cell‐T cell interaction. However, the role of the dendritic cells, and, indeed, of the antigen‐presenting step, has not yet been explored with respect to other T cell‐derived cytokines, in either a qualitative or relative fashion.

In this study, therefore, we have examined the comparative role of different antigen‐presenting cells (APC) as inducers of T cell cytokine release in allogeneic responses. We have confirmed that dendritic cells are the most effective inducers for IL 2 and have shown that this is true not only in primary alloresponses, but also in alloresponder T cells maintained for extended periods and then rechallenged. Dendritic cells were also the most potent inducers of IL 3 and interferon‐γ (IFN‐γ) in primary cultures. No IL 4 was demonstrable irrespective of the type of presenting cells used, and both tissue macrophages and dendritic cells can induce synthesis of IL 6. Likewise, in secondary alloresponses both dendritic cells and to a lesser extent tissue macrophages induce release of IL 3, no IL 4 is detectable, and activated macrophages and B cells raise IFN‐γ levels in the supernatants albeit to a lower concentration than that seen when dendritic cells are used as stimulators. The results were similar in the tertiary alloresponse except that (a) IL 4 was now detectable in the supernatants but only where dendritic cells had been used as APC, and (b) both resting and activated macrophages induced IL 2 and IFN‐γ. By the eighth cycle of allostimulation there is negligible IL 2. Dendritic cells, tissue macrophages and activated B cells constitute a hierarchy of APC for IL 3, IFN‐γ and IL 4. These findings therefore demonstrate the role of dendritic cells as potent in vitro inducers of IL 3, IL 4 and IFN‐γ synthesis as well as of IL 2.
Rejection of bone marrow cell allografts by natural killer cell subsets: 5E6+ cell specificity for Hh‐1 determinant 2 shared by H‐2d and H‐2f
Authors: Sentman Charles L. , Kumar Vinay , Bennett Michael
Published: March 2007

The 5E6 antigen, defined by anti‐5E6 mAb, is expressed on one‐half of murine natural killer (NK) cells, and we have previously demonstrated (C. L. Sentman et al., J. Exp. Med. 1989. 170: 1991) that 5E6+ NK cells are necessary for the rejection of BALB/c (Hh‐1d) but not C567BL/6 (Hh‐1b) bone marrow cells (BMC). In experiments described here, we have characterized the specificity of 5E6+ and 5E6− NK cell subsets for hemopoietic histocompatibility‐1 (Hh‐1) antigens. Prospective recipient mice were treated with anti‐5E6 mAb and challenged with BMC from a variety of donors. In addition, H‐2d/Hh‐1d C.B‐17 scid 5E6+ or 5E6− NK cells were adoptively transferred into irradiated, NK cell‐depleted hosts and challenged with H‐2b/Hh‐1b BMC. The data indicate that the 5E6+ NK cells are necessary for the rejection of only those BMC that express the Hh‐1 determinant 2 shared by H‐2d and H‐2f haplotypes of strains BALB/c (d), A.Ca (f), and B10.M (f). No reactivity to other Hh‐1 antigens resides in the 5E6+ population. In contrast, the ability of NK cells to lyse H‐2d or H‐2b tumor cells was independent of 5E6 expression. These results suggest that the 5E6 molecule is likely to be important in the specific recognition and rejection of BMC that express Hh‐1 determinant 2, and is probably not involved in recognition of „tumor target cell structures”︁.
Regulation of immune response to inhaled antigen by alveolar macrophages: differential effects of in vivo alveolar macrophage elimination on the induction of tolerance vs. immunity
Authors: Thepen Theo , McMenamin Christine , Oliver Jane , Kraal Georg , Holt Patrick G.
Published: March 2007

A single intratracheal instillation of liposomes containing dichloro methylene diphosphonate into rats eliminated > 80% of the alveolar macrophage (AMϕ) population, and the population was not significantly renewed during the ensuing week. AMϕ depletion markedly increased local antibody production in the lung wall in pre‐primed animals exposed to antigen aerosols. However, AMϕ depletion did not affect the normal development of protective tolerance (particularly in the IgE antibody class) to inhaled antigen in immunologically naive rats. These results are discussed in relation to regional control of immune responses in the upper vs. the lower respiratory tract.
Correction of antigen‐specific T cell defects in aged murine gut‐associated lymphoid tissues an immune intervention by combined adoptive transfer of an antigen‐specific immunoregulatory CD4 T cell subset and interleukin 2 administration
Authors: Kawanishi Hidenori , Ajitsu Shin
Published: March 2007

Gut‐associated lymphoid tissues (GALT) from aged mice enterically immunized with Mycobacterium paratuberculosis protoplasmic antigen show hyperreactive humoral immune responses; this hyperresponsiveness can be corrected to a considerable extent, but not entirely, by systemic administration of interleukin 2 (IL 2) alone. The aim of the present study was to determine further whether the hyperreactivity in the antigen‐specific humoral immune responses in aged GALT could be fully restored by adoptive transfer of in vitro expanded antigen‐specific IL 2‐dependent helper (CD4+VV−) T cells from GALT in conjunction with recombinant IL 2 administration. The results show that the age‐associated hyperresponsiveness in gut mucosal antigen‐specific humoral immune responses can be entirely corrected by adoptive transfer with the antigen‐specific GALT T helper cells together with in vivo IL 2 adminstration. The mechanism of this restoration involves reversal of the decline in antigen‐specific CD8+ suppressor T cell functions in aged GALT.
Homing receptors reexamined: mouse LECAM‐1 (MEL‐14 antigen) is involved in lymphocyte migration into gut‐associated lymphoid tissue
Authors: Hamann Alf , Jablonski‐Westrich Dorothee , Jonas Petra , Thiele Heinz‐Günter
Published: March 2007

Specific recognition molecules („homing receptors”︁) on lymphocytes are thought to direct selective entry of cells into different organs. The lectin‐related cell adhesion molecule LECAM‐1 has previously been supposed to mediate lymphocyte entry into peripheral lymph nodes and, partially, mesenteric nodes but not into Peyer's patches. Here we present evidence that in vivo the molecule is also implicated in homing of mouse lymphocytes to Peyer's patches and may have a more general role as homing receptor for high endothelial venules‐bearing lymphoid tissue, but not for most non‐lymphoid tissue.
Identification of thymocyte precursors in murine fetal liver
Authors: Bell Sarah E. , Zamoyska Rose
Published: March 2007

We have isolated thymic stem cells from murine fetal liver using a combination of approaches including discontinuous albumin density gradient separation and anti‐heat‐stable antigen (HSA) antibody plus complement‐mediated lysis. A population of early precursors has been identified which comprises approximately 1% of fetal liver, and which has the capacity to reconstitute alymphoid lobes and give rise to the four thymic subpopulations defined by expression of the T cell differentiation antigens CD4 and CD8. These early precursors express CD44 (Pgp‐1), however, they are negative for expression of Thy‐1, CD4, CD8, CD3, membrane immunoglobulin and HSA cell surface molecules.
Resistance to Leishmania major infection correlates with the induction of nitric oxide synthase in murine macrophages
Authors: Liew Foo Y. , Li Yun , Moss David , Parkinson Clive , Rogers Mark V. , Moncada Salvador
Published: March 2007

Inbred strains of mice differ considerably in their innate resistance to leishmanial infection. BALB/c mice are highly susceptible to cutaneous leishmaniasis caused by Leishmania major, whereas CBA mice are resistant. We now show that this resistance correlates with the ability of macrophages to synthesize nitric oxide (NO) following activation with interferon‐γ or tumor necrosis factor α. Furthermore, the larger amounts of NO generated by resistant macrophages are related to higher levels of NO synthase activity, a difference which is not attributable to the number or the affinity of the receptors for interferon‐γ on these cells. The level of NO synthesis by activated macrophages was also correlated to the resistance in a number of other inbred mouse strains tested; macrophages from the resistant B10.S, C57BL and C3H mice produced significantly higher levels of NO than the macrophages from the susceptible BALB.b and DBA/2 mice.
Immunogenicity of multiple antigen peptides (MAP) containing T and B cell epitopes of the repeat region of the P. falciparum circumsporozoite protein
Authors: Munesinghe Dona Yamuna , Clavijo Pedro , Calle Mauricio Calvo , Nussenzweig Ruth S. , Nardin Elizabeth
Published: March 2007

The immunogenicity of multiple antigen peptides (MAP) constructs containing T and B cell epitopes of the repeat region of the P. falciparum circumsporozoite (CS) protein was examined in vitro, using a human T cell clone, and in vivo, using four different strains of mice. All the MAP constructs that contained the T cell epitope, (DPNANPNVDPNANPNV), stimulated proliferation and interferon‐γ production by a human T cell clone specific for this epitope which is located in the 5′ end of the repeat region of the P. falciparum CS protein. These human T cells did not recognize MAP that contained only the B cell epitope, (NANP)3, which is located in the 3′ repeat region. Optimal antibody responses were obtained in mice immunized with MAP containing four copies of tandemly arranged T and B cell epitopes, (TB)4. The murine immune response to the MAP constructs was genetically restricted. Mice of a high responder strain, C57BL, recognized both the 5′ and 3′ repeat sequences in the MAP as T, as well as B, cell epitopes and developed very high anti‐MAP and anti‐sporozoite antibody titers. A/J and C3H mice, which were intermediate responders, developed lower antibody titers which varied according to the orientation of the T vs. the B cell epitopes within the MAP constructs. BALB/c mice were nonresponders and did not develop antibodies following immunization with any of the MAP constructs containing the 5′ and 3′ repeats of the P. falciparum CS protein.
Immunoglobulin heavy chain cDNA from the teleost Atlantic cod (Gadus morhua L.): nucleotide sequences of secretory and membrane form show an unusual splicing pattern
Authors: Bengtén Eva , Leanderson Tomas , Pilström Lars
Published: March 2007

Rabbit antibodies to Atlantic cod (Gadus morhua L.) immunoglobulin were affinity purified and used to screen cDNA libraries from spleen and head kidney mRNA. cDNA clones for both the secretory and membrane‐bound heavy (H) chain were isolated, the nucleotide and deduced amino acid sequences of which are reported here. Comparisons of the cod secretory H chain amino acid sequence show 24%, 27%, 30% identity to the μ chain of Mus, Xenopus and Ictalurus, respectively. The highest degree of identity was observed in the CH4 domain. The cDNA encoding the transmembrane form shows a novel splicing pattern where the TM1 exon is spliced directly onto the CH3 domain and not to the CH4 domain as in other animal groups. Southern blot analyses with VH and C probes on genomic DNA from cod erythrocytes indicate that there is a unique C gene but several V genes in the cod immunoglobulin H chain locus.
Monoclonal antibodies raised against engineered soluble mouse T cell receptors and specific for Vα8‐, Vβ2‐ or Vβ10‐bearing T cells
Authors: Necker Antje , Rebaï Najet , Matthes Michaela , Jouvin‐Marche Evelyne , Cazenave Pierre‐André , Swarnworawong Pam , Palmer Ed , Robson MacDonald H. , Malissen Bernard
Published: March 2007

We have characterized a panel of monoclonal antibodies (mAb) produced by immunizing rats with two distinct soluble mouse α/β T cell receptor (TcR). Fifty mAb were found to react with the corresponding surface‐bound TcR. Such observations suggest that the soluble TcR molecules used as immunogen are folded in a conformation similar to the native structure. Furthermore, the binding to T cells of four antibodies was found to correlate with the expression of the Vα8, Vβ2 or Vβ10 gene segments. Finally, staining of T lymphocytes from various mouse strains suggests that (a) the two anti‐Vα8 antibodies recognize different epitopes, and each on only a fraction of Vα8+ cells; (b) the anti‐Vβ10 mAb identifies a Vβ10 polymorphism among mouse strains, and (c) T cells expressing the Vβ2 or Vβ10 gene segments are not subject to major clonal deletion events induced by the major histocompatibility complex class II and Mls products which were tested.
Is early repopulation of macrophage‐depleted lymph node independent of blood monocyte immigration?
Authors: Mebius Reina E. , Martens Gratia , Brevé John , Delemarre Frans G. A. , Kraal Georg
Published: March 2007

Popliteal lymph nodes (LN) of mice were depleted of their macrophage (Mϕ) populations in the subcapsular sinus and medulla by subcutaneous injection of dichloromethylene diphosphonate (Cl2MDP)‐containing liposomes into the footpads. Complete restoration of both Mϕ populations could be observed as late as 5 months after liposome administration. This relatively long repopulation time could be due to a depot of liposomes, directly killing all Mϕ precursors after extravasation into the interstitial tissue of the footpad. On the other hand, local interstitial precursors with very low turnover rates may have been depleted in the interstitial tissue of the hind leg. Therefore, two different types of experiments were performed; one in which Mϕ‐depleted LN were replaced by control LN at various time points after liposome treatment, and another whereby Mϕ‐depleted LN were transplanted into control animals.

When liposome‐treated, Mϕ‐depleted LN were transplanted into control animals, a complete restoration of both populations in the subcapsular sinus and medulla could be observed within 5 weeks. Control LN transplanted into a Cl2MDP‐liposome‐treated leg showed a rapid disappearance of Mϕ from the subcapsular sinus and medulla and these cell populations remained absent for at least 7–8 weeks after liposome treatment, when the first cells started to reappear. Complete repopulation of these areas by Mϕ took as long as 15 weeks. Using labeled liposomes the presence of a continuous liposome depot was found to be very unlikely.

These results suggest that the population of precursor cells that will give rise to Mϕ in the subcapsular sinus and medulla of a LN is probably contained within the interstitial tissue and is almost independent of precursor supply from the blood compartment.
The ligand recognized by ELAM‐1 on HL60 cells is not carried by N‐linked oligosaccharides
Authors: Leeuwenberg Jet F. M. , Tan Agnes , Jeunhomme Trudi M. A. A. , Ploegh Hidde L. , Buurman Wim A.
Published: March 2007

The sialyl Lewis‐x determinant is a ligand for ELAM‐1, a major adhesion molecule for HL60 cells and neutrophils. ELAM‐1 expression can selectively be induced on human umbilical vein endothelial cells (HUVEC) by tumor necrosis factor, interleukin 1 and lipopolysaccharide. The determinant sialyl Lewis‐x is found on both glycolipids as well as on glycoproteins. Using specific inhibitors of the biosynthesis of N‐linked glycosylated glycoproteins, we investigated whether N‐linked glycans or their modifications are involved in ELAM‐1‐dependent adhesion of HL60 cells to activated HUVEC. The inhibitors of glycoprotein processing N‐methyl‐deoxynojirimycin, 1‐deoxymannojirimycin and swainsonine did not affect ELAM‐1‐dependent adhesion. Complex‐type N‐linked glycans are not required for ELAM‐1‐mediated adhesion, and therefore the ligand for ELAM‐1 is most likely a glycolipid, or a glycoprotein carrying O‐linked oligosaccharides.
Internalization of glycosyl‐phosphatidylinositol (GPI)‐anchored lymphocyte proteins II. GPI‐anchored and transmembrane molecules internalize through distinct pathways
Authors: Bamezai Anil , Goldmacher Victor S. , Rock Kenneth L.
Published: March 2007

Ly‐6A.2 (T cell‐activating protein, TAP) and Thy‐1 are glycosyl‐phosphatidylinositol (GPI)‐anchored proteins expressed on the surface of murine T lymphocytes. We have found that Ly‐6A.2 (TAP) and Thy‐1 are internalized by T cells. In the present study we have investigated whether these GPI‐anchored proteins enter cells by endocytosis through coated pits. Two lines of evidence argue against the involvement of coated pits in the internalization of Ly‐6A.2 (TAP) and Thy‐1. First, drugs that effectively blocked the endocytosis of transferrin receptor and H‐2 class I molecules, (which are known to be internalized via coated pits) did not inhibit the internalization of the GPI‐anchored proteins. Second, in ultrastructural analyses, Ly‐6A2 (TAP) and Thy‐1, in contrast to the transferrin receptor, were rarely found in coated pits or vesicles. These observations suggest that the GPI‐anchored proteins on T lymphocytes are internalized by a distinct pathway that does not involve endocytosis through coated pits.
Specificity and T cell receptor β chain usage of a human collagen type II‐reactive T cell clone derived from a healthy individual
Authors: Yan Tan , Burkhardt Harald , Ritter Thomas , Bröker Barbara , Mann Karl Heinz , Bertling Wolf M. , Mark Klaus Von Der , Emmrich Frank
Published: March 2007

Collagen type II (CII) is a cartilage‐specific matrix compound well known as an inducer of an experimental. T cell‐dependent autoimmune arthritis, a disease which shows some similarities to human rheumatoid arthritis. Here we report on an HLA‐DR7‐restricted human CD4 T cell clone (TC9), which was isolated from a healthy donor and recognizes human CII. After screening CNBr fragments of CII and tryptic fragments derived thereof, the T cell epitope could be mapped to amino acid residues 271‐285 of the triple helical region of CII that are located within CNBr fragment 11 [α1 (II) CB11]. This epitope was confirmed by a synthetic peptide stimulatory for TC9. The T cell receptor β chain of TC9 was cloned using the polymerase chain reaction; it comprises Vβ6.7 and contains besides Jβ2.3 and Cβ2 an as yet undescribed sequence for the D segment.
Genetic correction of cultured T cells from an adenosine deaminase‐deficient patient: Characteristics of non‐transduced and transduced T cells
Authors: Braakman Eric , Van Beusechem Victor W. , Van Krimpen Brigitte A. , Fischer Alain , Bolhuis Reinder L. H. , Valerio Dinko
Published: March 2007

T lymphocytes derived from peripheral blood of a patient with adenosine deaminase (ADA) deficiency were expanded in vitro. The human ADA (hADA) gene was introduced into these replicating ADA− T cells with the use of an amphotropic recombinant retrovirus carrying the hADA gene. Subsequently, infected T cells were selected on the basis of their ADA expression, by exposure to a combination of the toxic agent xylofuranosyl‐adenine and the specific ADA inhibitor 2′‐deoxycoformycin. CD4+ and CD8+ T cells could be infected and selected with equal efficiencies. The genetically modified T cells were shown to contain intact copies of the provirus and to express normal levels of hADA. As expected, uninfected T cells from the ADA‐deficient patient displayed an increased sensitivity to 2′‐deoxyadenosine. Following genetic modification, however, this sensitivity was restored to normal levels in both CD4+ and CD8+ T cells. The introduction of the hADA gene into the genome of the in vitro expanded T cells did not alter their phenotype, proliferative capacity or cytotoxic potential. These characteristics were identical to those of T cells derived from healthy individuals. These findings are of critical importance for the clinical application of hADA gene‐transducted T cells.
Thymic nurse cell lymphocytes react against self major histocompatibility complex
Authors: Penninger Josef , Wick Georg
Published: March 2007

It has been postulated that thymic nurse cells (TNC), lymphoid‐epithelial complexes composed of thymocytes enclosed within major histocompatibility complex (MHC) class I+ and class II+ cortical epithelial cells, may provide an optimal microenvironment for the process of T cell selection. By transplanting single TNC in the avian chorionallantoic membrane assay we demonstrate that a significant portion of intra‐TNC lymphocytes (TNC‐L) possess reactivity against self‐MHC molecules. The frequency of these autoreactive cells among TNC‐L exceeds by far that of thymocytes or peripheral blood lymphocytes of the same donor. These results indicate that TNC‐L constitute a T cell population enriched for self‐MHC reactivity, i.e. cells that have undergone positive selection, but not yet deletion and/or deactivation.
Novel excision products of T cell receptor γ gene rearrangements and developmental stage specificity implied by the frequency of nucleotide insertions at signal joints
Authors: Iwasato Takuji , Yamagishi Hideo
Published: March 2007

We have cloned circular DNA excised by T cell receptor (TcR)γ1, γ2 and γ3 gene rearrangements in fetal and adult mouse thymocytes. Circular DNA contained a signal joint reciprocal to the genomic V‐J coding joint. Although signal joints without nucleotide insertions are common in immunoglobulin (Ig) and TcR gene rearrangements, the signal joint of γ found in adult thymocytes contained non‐germ‐line element (N) insertions at high frequency, while no insertions were found in fetal thymocytes. Thus developmental stage specificity of TcR γ gene rearrangements is faithfully reflected on the signal joint of excision products. In addition, examination of γ gene excision products revealed circular DNA products of TcR γ‐α transrearrangements, but no evidence for Vγ gene replacement in a rearranged segment.
Studies on the induction of anterior chamber‐associated immune deviation (ACAID) III. Induction of ACAID depends upon intraocular transforming growth factor‐β
Authors: Wilbanks Garth A. , Mammolenti Michele , Streilein J. Wayne
Published: March 2007

Delayed hypersensitivity (DH), the prototypical form of cell‐mediated immune responsiveness, is mediated with the participation of considerable nonspecific inflammation which necessarily disrupts the anatomic integrity of involved and adjacent tissues. Damage of this type is of minor consequence to many visceral and cutaneous organs, but is of devastating consequence for organs such as the eye and the brain. At least in the case of the eye, the organ is remarkably adept at regulating the immune system's ability to respond to intraocular antigens by selectively down‐regulating both the induction and expression of delayed hypersensitivity while leaving other effector modalities intact. This ability of the eye to selectivity down‐regulate systemic DH responses to intracamerally inoculated antigens is known as anterior chamber‐associated immune deviation (ACAID) and is mediated in part by antigen‐specific regulatory T cells. Recent work suggests that macrophages (MΦ) that reside in the iris and ciliary body can migrate out of an antigen‐bearing eye and activate regulatory T cells within the spleen. In an effort to understand the mechanism by which intraocular MΦ interact with antigen in the anterior chamber of the eye (AC) and subsequently induce splenic regulatory cells in ACAID, we have investigated what role, if any, the AC microenvironment itself plays in ACAID induction. The results reveal that CD45− parenchymal iris/ciliary cells secrete a soluble factor(s) locally and into the aqueous humor which endows resident, mature MΦ with ACAID‐inducing capabilities. Mice receiving infusions of these altered, antigen‐pulsed MΦ are incapable of mounting a significant DH response following immunization with antigen in adjuvant. Importantly, the ACAID‐inducing effect is achieved when conventional, extraocular MΦ are exposed in vitro to a soluble factor present in aqueous humor or culture SN from iris and ciliary body cells. Further investigations into the identity of this factor reveal it to be transforming growth factor‐β (TGF‐β). The role of TGF‐β in the generation of ACAID, as well as the implications of these findings to an understanding of immunologic privilege in general, are discussed.
Augmentation of major histocompatibility complex class I and ICAM‐1 expression on glial cells following measles virus infection: Evidence for the role of type‐1 interferon
Authors: Kraus Elmar , Schneider‐Schaulies Sibylle , Miyasaka Masayuki , Tamatani Takuya , Sedgwick Jonathon
Published: March 2007

An intracellular staining procedure for the cytoskeletal marker, glial fibrillary acidic protein of astrocytes, has been developed which allows flow cytometric phenotyping of astrocytes within complex mixtures of glial cells. Employing this technique, we show here that measles virus infection of rat mixed glial cell cultures results in a rapid augmentation of major histocompatibility complex (MHC) class I and ICAM‐1 on the majority of astrocytes in culture. MHC class I levels are increased on macrophages/microglia but ICAM‐1 expression is not normally affected on this cell type. Some MHC class II induction is also observed after virus infection but only on astrocytes. A type‐I interferon (IFN)‐inducible protein, Mx, was identified in cultured glial cells after infection. Qualitatively comparable MHC class I and ICAM‐1 enhancement after addition of type‐I IFN, supports the conclusion that this cytokine(s) released as a result of virus infection, is responsible for alterations in the expression of molecules on glial cells, that are involved in T cell recognition. Astrocytes after viral infection were more susceptible to alloantigen‐specific cytotoxic T lymphocytes and cytotoxic T lymphocyte activity was substantially reduced in the presence of mAb specific for MHC class I, ICAM‐1 and LFA‐1 but not MHC class II. The relevance of these findings to T cell recognition of virus‐infected cells in the central nervous system is discussed.
Acute human vs. mouse graft vs. host disease in normal and immunodeficient mice
Authors: Huppes Wim , Geus Bernard De , Zurcher Chris , Van Bekkum Dirk W.
Published: March 2007

Recent reports of persistent engraftment of human lymphocytes and myeloid cells in hereditary immunodeficient severe combined immunodeficient mice (SCID) and beige athymic nude X‐linked immunodeficiency (Bg/Nu/XID) mice have raised the question of why attempts to graft human cells into artificially immunosuppressed normal mice have failed so far. In the present study we provide evidence that this difference is due to the absence of natural antibodies in the mutant mice. We demonstrate that human PBL can be grafted in normal mice immunosuppressed by heavy doses of total body irradiation, provided the transplant is performed when the recipients lack natural antibodies in their serum, e.g. as in newborn normal mice, in mice treated with anti‐mouse IgM antibody from birth, and in 3‐week‐old B cell‐deficient CBA/N mice. In all cases, large numbers of human PBL were required.

Under these conditions an acute and fatal graft vs. host disease (GVHD) developed in the recipients, regardless of whether these were artificially immunosuppressed or hereditary immunodeficient. The clinical manifestations and the histopathology of this xenogeneic acute GVHD are quite different from those of allogeneic GVHD. The former is primarily confined to the hematolymphoid tissues and locations close to accumulations of proliferating lymphoblasts, such as the peritoneal cavity in case of i.p. transplantation. The discordant xenogeneic GVHD is induced by human T lymphocytes and can be abrogated by treatment with anti‐human T cell serum.
Tolerance of self induced in thymus organ culture
Authors: Robertson Katherine , Simon Katharina , Schneider Susanne , Timms Emma , Mitchison Avrion
Published: March 2007

F liver protein occurs in serum at low concentration, and therefore induces tolerance of self only in T cells. T cells which mature in cultured thymus lobes in the absence of this protein become reactive towards it but can be prevented from doing so by exposure to the protein while in culture. The threshold of tolerance induction for this soluble antigen is estimated in this way at ∼ 1 μg/ml, which is slightly less than the threshold of response of primed T cells in a proliferation assay. Freshly isolated thymocytes do not display reactivity to self‐F protein, indicating that T cells normally become tolerant while still within the thymus.
Induction of experimental autoimmune encephalomyelitis by native myelin basic protein‐activated T lymphocyte lines
Authors: Friedman Aharon , Reshef Tamara , Cohen Irun R.
Published: March 2007

Self antigens bind to MHC class II molecules in vivo. The capacity of class II molecules to bind native self antigen, namely antigen that has not been processed through an endososomal pathway, could increase susceptibility to autoimmune diseases if recognized by autoreactive T lymphocytes. To confirm this prediction we designed experiments to show that: (a) native myelin basic protein (BP) activates encephalitogenic T lymphocyte lines, and (b) these activated lines cause experimental autoimmune encephalomyelitis (EAE) in naive rats. We show that two encephalitogenic T lymphocytes lines, LEW and BN, are activated by native BP in the presence of paraformaldehyde pre‐fixed antigen‐presenting cells (APC). The degree of activation, as measured by T lymphocyte proliferation, was equal to that obtained in response to BP processed by APC prior to paraformaldehyde fixing. The response to native BP was confirmed by demonstrating insensitivity to the presence of the lysosomotropic agent chloroquine or protease inhibitors. Activation of T lymphocytes by BP required the presence of syngeneic APC. The activated T lymphocyte lines were injected into naive recipient rats that developed EAE within 5 days. Both disease incidence and severity were equal to that observed in rats that were treated with T lymphocytes activated by processed BP. Hence, at least in EAE, the risk of an autoimmune event could be precipitated by a complex of native antigen and class II molecules on cells that do not possess an endososomal pathway for antigen processing and presentation.
Hierarchy of responsiveness in vivo and in vitro among T cells expressing distinct MIs‐1a‐reactive Vβ domains
Authors: Waanders Gary A. , McDonald H. Robson
Published: March 2007

In this report we have compared responses of T cells bearing different T cell receptor (TcR) Vβ domains, including Vβ6, 7, 8.1 and 9, to the minor lymphocyte stimulatory locus (MIs‐1a). The virtually complete deletion of peripheral T cells using these TcR Vβ domains in mice expressing MIs‐1a verified their MIs‐1a reactivity. However, varied responses among these cells following stimulation with MIs‐1a in vitro suggested a hierarchy of TcR reactivities for MIs‐1a with Vβ6 and Vβ9 > Vβ8.1 » Vβ7. This hierarchy was further supported by in vivo studies which showed that Vβ6+CD4+ cells dominated responses to MIs‐1a.
Phenotype, activation and lymphokine secretion by γ/δ T lymphocytes from schistosomiasis and carcinoma of the urinary bladder
Authors: Raziuddin Syed , Shetty Sugandh , Ibrahim Ahmed
Published: March 2007

In humans the majority of the CD3+ T cells usually express an α/ß T cell receptor (TcR) and a minority express a γ/δ TcR. The CD3+ TcR α/ß and CD3+ TcR γ/δ cells from blood of the patients with schistosomiasis with carcinoma of the urinary bladder (SCB) were analyzed for phenotype, activation, secretion of interleukin 2 (IL2), B cell growth factor (BCGF) and B cell differentiation factor (BCGF), as well as for autologous (AMLR) and allogeneic (MLR) mixed lymphocyte reaction. Patients with SCB had a highly increased percentage of CD3+ TcRγ/δ and a decreased percentage of CD3+ TcR γ/ß T cells in their circulation. These CD3+ TcR γ/δ T cells expressed the CD25 (IL 2 receptor), CD38, CD71 (transferrin receptor) and HLA‐DR activation antigens at a higher intensity after in vitro stimulation with recombinant IL 2, phytohemagglutinin and soluble egg antigen (from Schistosoma haematobium). The SCB patients' CD3+ TcR γ/δ T cells were highly deficient in secretion of IL 2 but produced highly elevated levels of BCGF and BCDF. On the contrary, both BCGF and BCDF activities of the CD3+ TcR α/β T cells were decreased. Moreover, CD3+ TcR γ/δ T cells demonstrated highly deficient AMLR and MLR activity. These observations suggest a possible role of CD3+ TcR γ/δ T cells in the immune response and the disease pathogenesis in human schistosomiasis infections.
Increased frequency of 2,4,6‐trinitrophenyl (TNP)‐specific, H‐2b‐restricted cytotoxic T lymphocyte precursors in transgenic mice expressing a T cell receptor β chain gene from an H‐2b‐restricted, TNP‐specific cytolytic T cell clone
Authors: Iglesias Antonio , Hansen‐Hagge Thomas , von Bonin Arne , Weltzien Hans Ulrich
Published: March 2007

T cell antigen receptors (TcR) expressing Vα10/JαBBM142 genes in association with β chains containing Jβ2.6 elements were found to be predominant among 2,4,6‐trinitrophenyl (TNP)‐specific, H‐2b‐restricted cytolytic T cell lines (CTL). To assess the relative contribution of the TcR β chain to the TNP specificity as well as to the selection of the respective TcR α chain elements we generated transgenic mice expressing the TcR β chain of the H‐2b/TNP‐specific CTL clone BT7.4.1. The TcR of this clone does not belong to the type predominant among H‐2b/TNP‐specific CTL, as it consists of an α chain encoded by a Vα8/JαDO gene rearrangement and a Vβ2/Jβ1.1‐containing β chain. In the transgenic mice almost all T cells exclusively express the transgenic Vβ2 gene, as a result of allelic exclusion. TNP‐specific, H‐2b‐restricted precursors were found at 7‐ to 8‐fold higher frequency in these mice as compared with non‐trangenic littermates. In H‐2b/d heterozygous transgenic mice, an increased frequency of TNP‐specific precursors was found only in H‐2b, but not in H‐2d‐restricted CTL. Analysis of H‐2b/TNP‐specific CTL lines derived from Vβ2‐transgenic mice indicated a preferential association of the transgenic TcR β chain with endogenous α chains encoded by Vα8 and JαBBM142 genes. This suggests that the hapten TNP is recognized like typical peptide antigens by combinatorial TcR α and β contact sites.
Genesis of host IgE competence: perinatal IgE tolerance induced by IgE processed and presented by IgE Fc receptor (CD23)‐bearing B cells
Authors: Chen Swey‐Shen Alex
Published: March 2007

A murine model for studying life‐long IgE tolerance was previously developed in this laboratory by perinatal IgE injection into neonates. Herein, we demonstrated that normal and immortal CD23+ B cell lines presented processed IgE via CD23‐mediated endocytic pathway and triggered perinatal IgE tolerance. The observations were as followed: (a) CD23 on normal B cells or B cell hybridomas mediated IgE‐dependent perinatal IgE tolerance and total IgE deficiency; and lack of either antigen‐specific IgE or total IgE did not correlate with elevated levels of autologous anti‐IgE in individual mice; (b) IgE tolerance‐inducing capacity of CD23+ B cell hybridomas was augmented by treatment with antigen‐IgE complexes orinterleukin 4, and significantly inhibited by anti‐CD23 prior to IgE pulsing; (c) antigen‐IgE complexes were endocytosed and degraded in acid hydrolases‐containing vesicles; and IgE tolerance was abrogated by treating IgE‐pulsed 17A11 at 4°C or 20°C followed immediately by fixation, and by treating IgE‐pulsed 17A11 with metabolic inhibitors that elevated intracellular pH of the endocytic vesicles. In conclusion, this study suggested that one pivotal step of genetic control of IgE responses may be exercised at the early developmental stage of T cells of the IgE lineage, and that CD23 may facilitate capture of endogenously secreted IgE, and mediate endocytic processing and presentation of self IgE epitope(s), and thus contribute to the genesis of host IgE competence.
Regulated expression on human macrophages of endoglin, an Arg‐Gly‐Asp‐containing surface antigen
Authors: Lastres Pedro , Bellon Teresa , Cabañas Carlos , Sanchez‐Madrid Francisco , Acevedo Agustin , Gougos Anne , Letarte Michelle , Bernabeu Carmelo
Published: March 2007

Endoglin is an endothelial homodimeric membrane antigen containing the tripeptide arginine‐glycine‐aspartic acid (RGD), which is a recognition motif for adhesion receptors of the integrin family. We have investigated the expression of endoglin by monocyte/macrophage cells from different tissue compartments and at different stages of cell differentiation. Although endoglin is absent from peripheral blood monocytes, it is expressed by in vitro differentiated monocytes as determined by flow cytometry using the endoglin‐specific monoclonal antibody 44G4 and 8E11. Furthermore, Northern blot analyses revealed a correlation between the presence of endoglin mRNA and the surface expression of the antigen by in vitro differentiated monocytes. Immunostaining of frozen tissue sections with the 8E11 monoclonal antibody demonstrated the presence of endoglin not only in the endothelium of all the tissues studied, but also on the interstitial macrophages present in the red pulp of the spleen. Using as a model of macrophage differentiation monocytic cell lines treated with phorbol esters, we found that the reactivity of the 8E11 monoclonal antibody is greatly increased on U‐937 and HL‐60 cells during their PMA‐induced differentiation. These findings clearly demonstrate for the first time the regulated expression of the putative adhesion molecule endoglin by macrophages.
Homologous collagen‐induced arthritis in ratg and mice are associated with structurally different major histocompatibility complex DQ‐like molecules
Authors: Holmdahl Rikard , Vingsbo Carina , Hedrich Hans , Karlsson Mikael , Kvick Christina , Goldschmidt Tom J. , Gustafsson Kenth
Published: March 2007

Collagen‐induced arthritis (CIA) in rats, induced with homologous type II collagen (CII), is a genetically more restricted disease and has better resemblance to rheumatoid arthritis by its chronic disease course, than CIA induced with heterologous CII. The DA strain is highly susceptible to CIA induced with homologous CII, while the Lewis strain is resistant. (DAxLew)F1 is susceptible and backcrossing to Lewis reveals a close, but not complete, association of both arthritis and CII responsiveness to the RT1a haplotype. Analyses of congenic strains on DA and Lewis backgrounds suggest that expression of a major histocompatibility complex class II Ba molecule, encoded from the RT1Ba locus, is associated with arthritis susceptibility and CII responsiveness. The second exons coding for the first domains of the α and β chains of both the RT1a and RT11 haplotypes were sequenced and the deduced amino acid sequences compared with the corresponding molecule associated with susceptibility to CIA in the mouse (H‐2 Aq). The sequences of the respective alleles revealed no obvious structural homology explaining the extensive similarities in the development of chronic autoimmune arthritis. Instead, this finding implies that different trimolecular constituents (i.e. class II,T cell receptor, and CII peptides) may yield an antigen presentation event that is able to trigger a similar autoaggressiveness in the two rodent species.