Compensation between Vav-1 and Vav-2 in B cell development and antigen receptor signaling

Vav-1 and Vav-2 are closely related Dbl-homology GTP exchange factors (GEFs) for Rho GTPases. Mutation of Vav-1 disrupts T cell development and T cell antigen receptor-induced activation, but has comparatively little effect on B cells. We found that combined deletion of both Vav-1 and Vav-2 in mice resulted in a marked reduction in mature B lymphocyte numbers. Vav-1(-/-)Vav-2(-/-) B cells were unresponsive to B cell antigen receptor (BCR)-driven proliferation in vitro and to thymus-independent antigen in vivo. BCR-stimulated intracellular calcium mobilization was greatly impaired in Vav-1(-/-)Vav-2(-/-) B cells. These findings establish a role for Vav-2 in BCR calcium signaling and reveal that the Vav family of GEFs is critical to B cell development and function.     

Signal transduction through the human IL-2 receptor beta-chain expressed in IL-6-dependent mouse B cell hybridoma

Non-covalent association between at least two polypeptides, alpha (p55) and beta (p70), yields a high-affinity interleukin 2 receptor (IL-2R). Recent findings suggest that the beta-chain can mediate IL-2 signals on its own, while the alpha-chain is not involved in IL-2 signal transduction. To study the role of the beta-chain, directly, we transfected with the human IL-2R beta-chain cDNA a murine IL-6-dependent B cell hybridoma, F12-28, which originally did not express IL-2R. We established a stable transformant, beta E12, expressing the beta-chain (Kd = 1300 pM, 3000 sites/cell) in the absence of any detectable alpha-chain. We showed that (i) beta E12 manifested the intermediate affinity IL-2 binding, which was completely blocked with anti-human beta-chain antibody (Mik-beta 1); (ii) beta E12 acquired an ability to proliferate in response to IL-2 (greater than 0.1 nM) in a dose-dependent manner. These results clearly demonstrate that the beta-chain itself is directly involved in IL-2 signal transduction in the absence of the alpha-chain. Our results also suggest that a certain IL-6-dependent B cell line possesses cellular components) capable of transducing IL-2 signals.     

Signal transduction from the B cell antigen-receptor.

Ligation of the B cell antigen-receptor triggers an intricate maze of intercalated biochemical events that ultimately affect B cell biological responses. Recent advances have helped to connect many loose ends by identifying key adaptor proteins, such as BLNK/SLP-65, defining crucial roles for phosphatidylinositol-3-kinase and mapping pathways controlling the mitogen-activated protein kinases (ERK, JNK and p38).     

Signal transduction in B lymphocytes

We have examined the activity and intracellular compartmentalization of protein kinase C (PKC) following activation of human B lymphocytes by anti-human leukocyte antigen (HLA) class II antibodies. 12-O-Tetradecanoylphorbol 13-acetate (TPA) treatment increased membrane-associated PKC (between five and nine times greater than the control value) and decreased cytosolic PKC (between 70% and 100% of the control value). In contrast, anti-class II antibodies induce an activation of PKC which results either in an increase of cytosolic activity or membrane-bound activity without redistribution of cytosolic PKC. The effect of TPA and HLA class II molecules on total PKC activity was comparable: when TPA induced an increase of total PKC activity so did HLA class II molecules and when TPA did not, HLA class II molecules did not. Measurement on SDS PAGE of histone phosphorylation confirmed the above results of PKC activity. Taken together, our results suggest that PKC might be implicated in HLA class II-induced B lymphocyte activation.     

IL-2 infusion abrogates humoral immune responses in humans.

Although IL-2 infusion enhances cell-mediated cytotoxicity in patients with neoplastic disease, administration is paradoxically associated with a modest fall in total serum IgG and an increased risk of infection. We now show that the adverse effects of IL-2 infusion on the humoral immune system are substantial. Although IL-2 induces the B cell growth and differentiating factors IL-4 and IL-6, infusion abrogates primary antibody responses entirely and reduces secondary antibody responses 50-fold following antigen challenge. There is no evidence of the generation of cells with suppressive activity on B cells but IL-2 increases the ratio of circulating virgin:memory cells. These results may help to explain the increased rate of bacterial infection in patients receiving IL-2. As IL-2 plays a central role in the generation of an immune response, the finding that it is also sufficiently immunosuppressive to inhibit primary- and secondary-type antibody responses suggests that exploration of the underlying mechanisms may provide insights into immune system homeostasis and may offer new approaches to therapeutic immunosuppression.     

HIV-2 DNA疫苗免疫小鼠的免疫反应观察

目的：用构建的HIV－2外膜蛋白gp105和核心蛋白gag基因的DNA疫苗免疫小鼠，评价其免疫效果。方法：将HIV－2型gp 105和gag基因克隆到表达载体pIRES1neo中，构建重组DNA疫苗质粒。间接免疫荧光试验证明，构建的DNA疫苗在真核细胞中表达了gp105或／和gag.构建的疫苗免疫小鼠后，用流式细胞仪测定CD4^ 、CD8^ T淋巴细胞亚类数，并用HIV－2抗体ELISA检测试剂盒检测免疫鼠血清中抗HIV－2抗体水平。结果：构建了3种HIV－2 DNA疫苗pIRES1gag、pIRSE1gp105和pIRES1gag-gp105，转染BHK细胞后均能表达抗原蛋白，免疫小鼠后可有效地刺激淋巴细胞增殖并诱导产生抗HIV－2特异性抗体，其中pIRES1gag-gp105免疫鼠中，淋巴细胞增殖最显著，而pIRES1gp105免疫鼠中HIV－2特异性抗体水平最高。结论：构建的DNA疫苗均能诱导机体产生免疫反应，其中pIRES1gp105诱导的体液免疫应答最显著，而pIRES1gag-gp105 诱导的细胞免疫响应最显著。     

Complement and the regulation of humoral immune responses

The complement component C3 is involved in the successful induction of a normal antibody response, but its mode of action is unclear. From studies with complement deficient guinea pigs Erik Böttger and Dieter-Suermann suggest that failure to stimulate antibody generation is the result of C3 depletion, rather than the suppression of activation by C3 fragments. In addition, they propose that complement may have a second regulatory function, being indirectly or directly involved in the suppression of the polyclonal activation of lymphocytes.     

The architects of B and T cell immune responses

Published work links adult lymphoid tissue-inducer cells (LTi) with T cell-dependent antibody responses. In this issue of Immunity, Tsuji et al. (2008) associate LTi with T cell-independent IgA antibody responses in the gut.     

Affinity maturation and hypermutation in a simulation of the humoral immune response

By experimenting with a cellular automaton model of the immune system, we have reproduced affinity maturation of the antibody response, a somatic adaptation to a changing environment. The simulation allowed the isolation of a number of variables, e.g. the fraction of repertoire available, the magnitude of the change in affinity with mutation, the mutation frequency and its focus on the complementarity-determining regions (CDR) of the antibody. Multiple series of immunizations were run in machina where the contribution of each variable was evaluated against the maturation observed. We found that hypermutation is not necessary for affinity maturation if the repertoire of B cell specificities is sufficiently complete, but is essential when the B cell diversity is limited (which happens to be the case in vivo), as it fills the holes in the repertoire and allows selection by antigen. Maturation also depends on the magnitude of the change in affinity with mutation, and we supply some necessary limits on this parameter. For mutations confined to the CDR, the most efficient maturation occurs at mutation rates of 0.2 per paratope and per cell division. When mutations also affect the framework regions, the peak of the most effective CDR mutation rate moves progressively to lower values. A most sensitive parameter is the speed of maturation, which reflects the rate of expansion of mutated clones. Comparing it with biological observations can help to discriminate between alternative hypotheses on the phenomena of hypermutation and affinity.     

Distinct roles for B7 costimulation in contact hypersensitivity and humoral immune responses to epicutaneous antigen

Productive interactions between B7-1 and B7-2 costimulatory molecules on dendritic cells (DC) and CD28 on T cells are thought to be critical for successful antigen presentation. Epicutaneous application of haptens induces both contact hypersensitivity (CHS), an inflammatory cutaneous response mediated by CD8⁺ T cells, and an anti-hapten antibody response mediated by CD4⁺ helper T cells. The role of B7 costimulation in the immune response to oxazolone (Ox) was analyzed using mice lacking either B7-1 (B7-1 − / −), B7-2 (B7-2 − / −), or both (Db − / −) of these costimulatory molecules. The absence of both B7-1 and B7-2 results in diminished CHS. This inhibition is largely overcome at higher hapten sensitizing doses indicating the presence of compensatory pathways. In contrast, anti-Ox IgG1 and IgG2a responses were not detected in the absence of both B7-1 and B7-2, even at high sensitizing doses, indicating an obligatory role of B7 costimulation in IgG class switching. B7-1 and B7-2 have overlapping functions in both CHS responses and anti-hapten response. B7-2 − / − mice demonstrated a modestly reduced CHS response only at very low doses of Ox (0.05 %), but responded normally at higher Ox doses, and B7-1 − / − mice had CHS responses indistinguishable from those of wild-type mice. Similarly, anti-Ox IgG responses were comparable in wild-type, B7-1 − / − and B7-2 − / − mice. Taken together, these studies reveal distinct roles for B7 costimulation in response to epicutaneous antigens with an obligatory role for IgG class switching and an important, but nonessential role for CHS responses.     

Signal transduction elements of the B cell antigen receptor and their role in immunodeficiencies

The primary function of B lymphocytes is to contribute to the elimination of foreign antigens by producing large amounts of soluble antibodies. The activation of B cells through their antigen receptor triggers a dynamic network of intracellular signaling proteins. The recent identification of the cytoplasmic adaptor protein SLP-65 (also called BLNK or BASH) provided insight in how the antigen receptor-regulated protein tyrosine kinases couple to downstream signaling cascades, including the mobilization of Ca2+ ions, activation of mitogen-activated kinases and reorganization of the cytoskeleton architecture. While these events have been mostly studied in mature B cells, it is now clear that the components of the antigen receptor and its downstream effector elements play also a central role during early and late B cell development, and in the apoptotic elimination of B cells with reactivity to self-antigens. Thus, genetic defects affecting the expression of antigen receptor subunits or its intracellular signaling proteins can interfere with B cell development and activation, and can cause severe antibody deficiencies in mouse and man. In this article I summarize our current picture of the B cell antigen receptor, how the extracellular signal is transported into the cell interior, and how dysregulation of these processes contribute to immune defects.     

Cellular and humoral immune responses in Campylobacter pylori-associated chronic gastritis

Gastric cellular and humoral immune response investigated by immunoperoxidase staining of 53 antral biopsies showed significant differences in Campylobacter pylori-associated gastritis as compared with non-bacterial chronic gastritis and normal controls. IgA, secretory component, and complement C3 coated bacilli were seen in all cases of active chronic gastritis. C3 was always associated with coating by IgA, IgM, or both, which were rarely seen in gastritis without polymorphonuclear neutrophil infiltration. Intraepithelial mononuclear cellular infiltration was seen in 18 of 26 cases of C. pylori-associated chronic gastritis. The intraepithelial mononuclear cells stained positively for T cells and histiocytes.     

Nutrient sensing,signal transduction and immune responses

Most cells in the body have a constant supply of nutrients, which are required to sustain cellular metabolism and functions. In contrast, cells of the immune system can encounter conditions with a limited nutrient supply during the course of an immune response. Cells of the immune system frequently operate in complex nutrient restricted microenvironments such as tumour or inflammatory sites. The concentrations of key nutrients such as glucose and certain amino acids, can be low at these sites, and this can have an impact upon immune cell function. Nutrient sufficiency is important to supply the metabolic and biosynthetic pathways of immune cells. In addition nutrients can also act as important cues that influence immunological signalling pathways to affect the function of immune cells. This review will describe the various nutrient sensing signalling pathways and discuss the evidence that nutrients are critical signals that shape immune responses.     

Nature of immune lymph node factors stimulating humoral and cellular immune responses

Institute of Immunology, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. D. Ado.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 107, No. 4, pp. 460–462, April, 1989.     

The rheumatoid synovial membrane participates in systemic anti-viral immune responses.

Sixteen patients with rheumatoid arthritis were systemically immunized with influenza virus vaccine and in vitro anti-influenza antibody responses by blood lymphocytes and lymphocytes isolated from synovectomy specimens were measured after in vitro challenge with this influenza antigen. Synovial lymphocytes from eight of these patients produced anti-viral antibody, thereby indicating that infiltrating lymphocytes participate in a systemic anti-viral immune response.