A role for mature B cells in bone marrow transplantation.

The proliferative potential of membrane Ig (mIg)-bearing B lymphocytes was assessed in an adoptive transfer system based on the use of non-inbred rabbits matched for major histocompatibility (MHC) antigens and mismatched for immunoglobulin (Ig) allotypes. Cell suspensions made from spleens (SP), mesenteric lymph nodes (LN), or bone marrow (BM) of allotype b4b5 rabbits were deprived of B cells with mIg of the b4 type by adherence to plastic dishes coated with affinity-purified anti-b4. When such b4-depleted cell populations were injected into newborn hosts of allotype b6b6, stable and lasting chimerism promptly resulted, in which donor-derived products were almost entirely of the b5 allotype. Chimeras formed by transfer of unfractionated cells from b4b5 donors, on the other hand, exhibited a predominance of the b4 allotype, as seen in the living donors. BM but not SP or LN contained precursors capable of differentiating into mIg+ lymphocytes in culture, but no evidence was obtained for engraftment and differentiation by such B-cell precursors or more primitive stem cells in vivo. These studies suggest a potentially significant role for mature B cells in reconstituting the immune system of human transplant recipients.     

A role for adhesion molecules in contact-dependent T help for B cells.

The role of cell contact in T-dependent B cell activation was examined. Small resting B cells from C57BL/6 mice were cultured with CBA-derived, non-alloreactive cloned T helper cells in anti-T cell receptor V beta 8-coated microwells. This induced polyclonal B cell activation to enter cell cycle (as measured by thymidine incorporation at 2 days) and to secrete immunoglobulin (as measured by an enzyme-linked immunoassay detecting high-rate Ig secretion at 5 days). The inclusion of monoclonal antibodies against LFA-1. ICAM-1 and CD4 in these cultures strongly inhibited antibody responses, although proliferative responses were only inhibited to about 50%. Inhibitory monoclonal antibodies did not significantly affect lipopolysaccharide-induced responses. T cell activation to interleukin (IL) 3 secretion, nor did they inhibit the formation of multicellular clusters containing T and B cells. There was no correlation between the level of expression of adhesion molecules by T cells and their ability to induce B cell responses. Anti-LFA-1 abrogated T-dependent responses to IL2 which were inducible after 2 days in culture, but did not inhibit the induction of this IL2 responsiveness. These results suggest that continued cell contact involving adhesion/accessory molecules induces B cells to proliferate and to respond to T cell lymphokines. A signaling role for cell interaction molecules on B cells is proposed, similar to the role of these and analogous molecules on T cells.     

A crucial role for macrophages in the pathology of K/B x N serum-induced arthritis

Autoantibodies in the form of immune complexes are known to be crucial mediators in initiating inflammation in a variety of autoimmune diseases. This has been well documented in the anti-collagen II antibody-induced arthritis animal model for a long time now. Recently, in the K/B x N mouse model (the F1 of the TCR-transgenic KRN and the diabetic NOD mice), anti-glucose-6-phosphate isomerase (GPI) autoantibodies have been shown to induce arthritis. Experimental work in the K/B x N model demonstrated key roles of autoantigenic immune complexes activating the alternative pathway of complement, the subsequent association with C5aR and Fc gammaRIII-mediated cell activation and production of the inflammatory cytokines IL-1 and TNF-alpha, finally leading to joint destruction. The presence of high amounts of inflammatory cytokines and matrix-degrading proteases at sites of inflammation obviously put the cytokine-producing macrophages as the next target for investigation in this model. Here, we show that mice depleted of macrophages by clodronate liposome treatment are completely resistant to K/B x N serum-induced arthritis. Reconstituting clodronate liposome-treated mice with macrophages from naive animals could reverse this resistance. Also, we found that deficiencies in the Wiskott-Aldrich syndrome protein and CD40, which are both implicated in macrophage activation, chemotaxis and phagocytosis, are not essential in serum-induced arthritis. Mast cell degranulation was seen in arthritogenic serum-treated mice even in the absence of macrophages, possibly suggesting that mast cell degranulation/activation acts hierarchically before macrophages in the inflammatory cascade of anti-GPI antibody-induced arthritis.     

Mast cells play a crucial role in Staphylococcus aureus peptidoglycan-induced diarrhea

Bacterium-induced diarrhea results in 2 to 2.5 million deaths in the world each year. The mechanism needs to be further understood. Staphylococcus aureus infection has a close relation with diarrhea; its cell wall component peptidoglycan (PGN) has strong biological activity on immune cells and possibly plays a role in S. aureus-induced diarrhea. The present study showed that oral PGN-induced diarrhea in mice in a dose-dependent manner. Intestinal epithelial cells absorbed PGN via the intracellular pathway. Intestinal mast cells were activated after PGN gavage. Toll-like receptor (TLR)2 expression was detected in mast cells in the intestine as well as in the murine mast cell line p815 cells. Blocking TLR2 or nucleotide-binding oligomerization domain (NOD)1 with related antibodies or RNA interference abolished PGN-induced p815 cell activation. The mast cell mediator histamine and serotonin had synergistic effects in PGN-induced diarrhea. In summary, oral PGN can induce diarrhea in mice, and TLR2 and NOD1 mediate the PGN-induced mast cell activation that plays a critical role in diarrhea induction. Blockade of TLR2 or NOD1 or treating mice with a mast cell stabilizer can efficiently inhibit PGN-induced-diarrhea, providing potential therapeutic significance.     

Possible immunoregulatory role for CD5 + B cells.

CD5 + B cells represent a subpopulation of B cells which have the characteristic of employing unmutated immunoglobulin variable region genes. These cells are found to be increased early in ontogeny. The percentage of CD5 + B cells is highest in the fetus and decreases after birth. The antibodies produced by CD5 + B cells are polyreactive and are the natural autoantibodies. These autoantibodies may not be pathogenic. CD5 + B cells are elevated in certain autoimmune disease states and are the malignant cell type in B-CLL, with a strong genetic component involved in determining elevated CD5 + B cell states. Elevated CD5 + B cells are found in immunodeficient states (young, aged, and autoimmune). CD5 + B cells may normally act as a first-line defense against invading foreign pathogens but are not involved in the specific immune response. There is some evidence, at least in newborns, that CD5 + B cells may affect the emerging B cell repertoire of conventional B cells via idiotype cascade. However, the action of CD5 + B cells in the newborn may be quite different than their activity in the adult. Nonimmunoglobulin-producing CD5 + B cells may be immunosuppressors. In this report, a unique subpopulation of CD5 + B cells was investigated. These cells were found only in the spleens of aged NZB mice. The CD5 + B cells were clonal and possessed extra chromosomes and did not appear to be producing antibodies. These cells were capable of rapid proliferation in unirradiated recipients. By taking advantage of this proliferative capability, the effect of exogenous clonal CD5 + B cells on recipient immune system was evaluated. Clonal CD5 + B cells from NZB mice were immunosuppressive and decreased the numbers of conventional B cells as well as the level of "natural antibodies." In summary, CD5 + B cells may play different roles in the immune system depending upon environment, age, and their differentiation state (i.e., proliferation versus antibody secretion). The natural antibody produced by CD5 + B cells may be involved in maintenance functions such as removal of dead cells and first-line defense mechanisms. In addition, CD5 + B cells may themselves regulate the immune system and produce a factor which is immunosuppressive. An understanding of the various functions of CD5 + B cells may elucidate fundamental immunoregulatory circuits.     

Essential role for B cells in transplantation tolerance

T lymphocytes are the primary targets of immunotherapy in clinical transplantation. However, B lymphocytes are detrimental to graft survival by virtue of their capacity to present antigen to T cells via the indirect pathway of allorecognition and the generation of donor specific alloantibody. Furthermore, the long-term survival of organ allografts remains challenged by chronic rejection, a process in which activated B cells have been found to play a significant role. Therefore, the achievement of transplantation tolerance will likely require induction of both T and B cell tolerance to alloantigens. Moreover, human and animal investigations have shown that subsets of B cells, Transitional and Regulatory, are inherently tolerogenic. Developing therapeutic strategies that exploit these populations may be key to achieving transplantation tolerance. In this review we describe the current evidence for the essential role of B cells in transplant tolerance and discuss emerging B cell directed strategies to achieve allograft tolerance.     

A proposed role for the lutropin receptor in contact-inducible gonococcal invasion of Hec1B cells.

We previously reported the existence of a contact-inducible, enhanced invasion phenotype in the obligate human pathogen Neisseria gonorrhoeae. Our present studies showed that the ability of glutaraldehyde-fixed eucaryotic cells to convert gonococci (GC) to this invasive phenotype (Inv+) is limited to cells derived from reproductive tissues. We present evidence that GC recognize the lutropin receptor (LHr), which recognizes both luteinizing hormone and human chorionic gonadotropin (hCG), as the tissue-specific environmental signal that induces the conversion of GC to the Inv+ phenotype. By competitive binding studies, we showed that Inv+ GC bind to Hec1B cells, a human endometrial cell line, by a unique adhesin not present on noninduced GC and that this Inv+ GC-specific binding is completely blocked by the addition of hCG. We demonstrated that limiting the access of GC to LHr decreases the ability of the host cell to both convert GC to the Inv+ phenotype and serve as a target for Inv+ GC invasion. We propose a model of GC invasion of Hec1B cells in which the LHr plays a dual role both as an induction signal and as part of the internalization mechanism. This utilization of LHr could account for both the preponderance of complicated GC disease in women and the observed correlation of the disease with the onset of menses.     

Dendritic cells have a crucial role in the production of cytokines in mesenteric lymph nodes of B10.BR mice infected with Trichuris muris

Dendritic cells bridge innate and adaptive immunity and establish protective immunity to pathogens. Protection against the murine nematode parasite Trichuris muris depends on the T helper 2 (Th2) response and requires the Th2 cytokines interleukin 4 (IL-4), IL-10, or IL-13. To examine if the Th2 response to T. muris infection is regulated by CD11c⁺B220⁻ dendritic cells in mesenteric lymph nodes, dendritic cell-enriched and dendritic cell-depleted fractions were obtained from mesenteric lymph node cells of T. muris-infected mice, and production of cytokines in cultures of these fractions was measured. At day 14 postinfection, no worm expulsion was observed, and high levels of interferon γ production occurred in dendritic cell-enriched fractions. Expulsion of worms occurred on days 20 and 25 postinfection, and IL-10 production was induced in dendritic cell-enriched fractions on these 2 days. No cytokine production was observed in mesenteric lymph node cells and dendritic cell-depleted fractions during T. muris infection. The occurrence of worm expulsion was consistent with IL-10 production in dendritic cell-enriched fractions. IL-10 inhibits Th1 cells and promotes the Th2 response, and results from this study suggest that CD11c⁺B220⁻ dendritic cells in the mesenteric lymph nodes are required for IL-10 production and the IL-10-dependent protective Th2 response.     

Early-appearing tumour-infiltrating natural killer cells play a crucial role in the generation of anti-tumour T lymphocytes.

Natural killer (NK) cells that infiltrated into the primary tumour site at an early stage of tumour development, were examined for their participation in the generation of anti-tumour cytotoxic T lymphocytes (CTL). NK cells, which were detected by anti-NK1.1 monoclonal antibody (mAb), increased in the peritoneal exudate cells (PEC) on days 3 and 7 after an intraperitoneal (i.p.) inoculation of syngeneic B16 melanoma cells. These tumour-infiltrating NK cells showed a high level of cytotoxic activity against NK-sensitive YAC-1 cells and an increased expression of interferon-gamma (IFN-gamma) mRNA and interleukin-2 (IL-2) mRNA. The in vivo depletion of NK cells with anti-NK1.1 mAb, prior to i.p. inoculation of B16 melanoma cells, resulted in an increased number of tumour cells in the PEC compared to NK cell non-depleted mice. Interestingly, the differences in tumour cell number between both groups were more prominent on days 7 and 14 than on day 3, which strongly suggested that early-infiltrating NK cells have a large influence on the subsequent anti-tumour response. The in vivo depletion of NK cells prior to immunization with melanoma cells abrogated the capacity of the spleen cells to generate CD8+ tumour-specific CTL after in vitro restimulation. This inability of generating anti-tumour CTL was partially restored by additional i.p. injections of recombinant IL-2 and/or IFN-gamma simultaneously with the immunization of melanoma cells. The in vitro depletion of NK cells prior to the in vitro restimulation with melanoma cells partially impaired the anti-tumour CTL generation from the spleen cells of the immunized mice. Lastly, the in vivo depletion of NK cells prior to immunization with melanoma cells abolished the protective immunity against melanoma cells at the rechallenge. Overall, these results indicate that early-appearing tumour-infiltrating NK cells not only participate in the anti-tumour early defence by themselves, but also play a crucial role in the generation of anti-tumour CTL.     

The CD19 molecule is crucial for MID-dependent activation of tonsillar B cells from children

The Moraxella immunoglobulin (Ig) D-binding protein (MID) induces a strong proliferative response in human peripheral blood IgD+ B cells from adults isolated by positive selection using anti-CD19-conjugated microbeads. Here, we show that tonsillar B cells from children isolated with positive selection are unable to respond to MID stimulation. The proliferative response was very low or absent at various concentrations of MID tested and at different time points analysed, whereas the MID response of tonsillar B cells from adults isolated with positive selection was considerably higher. Tonsillar B cells from children isolated with positive selection responded to formalin-fixed preparations of Moraxella catarrhalis and Staphylococcus aureus Cowan strain I. In comparison to cells isolated with positive selection, a much higher proliferative response was recorded in tonsillar B cells from children isolated with negative selection, indicating that occupation of the CD19 molecule (i.e. positive selection) inhibited the response. Indeed, the addition of anti-CD19 monoclonal antibodies (MoAb) to MID-activated tonsillar B cells from children isolated with negative selection strongly inhibited the proliferative response. In contrast, anti-CD21 MoAb at the same concentration did only show a minor inhibition on the MID-induced response. Pre-incubation of tonsillar B cells isolated from children with anti-CD19 or anti-CD21 MoAb did not affect the binding of biotin-conjugated MID as analysed by flow cytometry. These results suggest that MID-activated tonsillar B cells from children have a strong requirement for signalling through the CD19 molecule. Future experiments will further reveal the importance of CD19 and possibly other molecules for optimal activation of tonsillar B cells isolated from both children and adults.     

The role of B cells and autoantibodies in rheumatoid arthritis.

In this article, we will review B lymphocyte development and function, then discuss the role of B cells in RA, including immune complex formation; the K/BxN mouse model of RA; toll-like receptors; B cells as antigen presenting cells; germinal center-like structures in RA synovium; and influence on T cell activation, leukocyte infiltration, and angiogenesis. With regard to autoantibody production, we will focus on rheumatoid factor (RF) and anti-CCP antibodies, particularly mechanisms of their production; sensitivity and specificity in RA; and their roles as prognostic factors. Other autoantibodies will be discussed, as will treatment implications and future areas of investigation related to B cells and autoantibodies in RA.     

The role of B cells in acute and chronic infections.

The important role of B cells in protection against secondary viral infections has been recognized for a long time. Recent evidence suggests that B cells are also critically involved in protective immune reactions classically attributed to T cells. Specifically, antibodies have been documented to protect from many primary viral and parasitic infections and to be indispensable for the control of latent viral infections. Current vaccine strategies should take into account this pivotal role of antibodies.     

A dual role of activin A in regulating immunoglobulin production of B cells

Here, we report that activin A has a dual role in regulating Ig production of murine B cells. Activated B cells secrete activin activity by increasing activin A and decreasing follistatin expression. B cells also express type I and type II activin receptors, suggesting that they are targets of activin. Pretreatment of na?ve B cells with activin A and subsequent activation by LPS resulted in increased cell growth and IgG production. In contrast, no significant effect was observed when activin A was added to na?ve B cells simultaneously with LPS, indicating that activin A acts on resting but not activated B cells. In addition, activin A did not induce B cells to produce IgE, even when added prior to activation; however, in vivo antigen-specific IgE production was reduced significantly by neutralization of circulating activin A. These findings indicate that activin A plays an important role in Th2-mediated immune responses by enhancing antibody production through two distinct modes: acts directly on resting B cells to elicit full functions of activated B cells and acts indirectly on activated B cells through modulation of other immune cells.     

A role for the VLA-4 integrin in the activation of human memory B cells

It is generally recognized that activation through membrane effector molecules such as CD40 or the B cell receptor (BCR) is mandatory to allow B cells to proliferate and differentiate into antibody (Ab)-secreting cells in response to cytokines. We show here that purified tonsillar B cells can be stimulated directly by a cytokine combination to proliferate and secrete immunoglobulins when cultures are performed at high cell density. The contact-mediated activation of B cells in this experimental system is strongly inhibited both by anti-very late antigen (VLA)-4 monoclonal Ab and by a peptide containing the LDV sequence specifically recognized by the α4 integrin binding site. These reagents also significantly suppressed the B cell responses elicited by engagement of the BCR or CD40. Our data reveal that memory B cells but not virgin or germinal center B cells are sensitive to the direct stimulatory effect of cytokines in high-density cultures. Finally, we found that the dual expression of the α and β chains of VLA-4 is a distinctive feature of the memory B cell population. Collectively, our findings support the notion that VLA-4-dependent homotypic B cell interactions can mediate a co-stimulatory signal to human memory B cells and might participate in the B cell activation triggered through the BCR and CD40.