Unresponsiveness to Mlsa induced in newborn Mlsb mice by maternal preimmunization.

Female BALB/c (H-2d, Mlsb) mice alloimmunized prior to and during syngeneic pregnancy with DBA/2 (H-2d, Mlsa) splenocytes gave rise to offspring with severely reduced responsiveness in adult life to DBA/2 stimulation in vitro mixed lymphocyte cultures. The offspring of the hyperimmunized mothers were also tolerant to neonatal challenge with large numbers of DBA/2 splenocytes, which resulted in runting disease of control neonatal BALB/c mice. Both challenged and unchallenged offspring of the immunized BALB/c mothers were hyporesponsive to DBA/2 but both stimulated and responded to normal BALB/c lymphocytes, indicating alteration in their T-cell repertoire. There was no reduction in the V beta 6-positive thymocyte subpopulation in the challenged or unchallenged offspring of the alloimmunized BALB/c mothers compared to normal controls, suggesting that hyporesponsiveness to DBA/2 is not due to thymic deletion of Mlsa-responsive clones. Examination of the T-cell subset composition of the hydrocortisone-resistant thymocytes and peripheral lymphocytes of the challenged and unchallenged groups of experimental mice revealed large increases in the percentage of Lyt-2+ T cells, sometimes accompanied by a decrease in the L3T4+ T-cell subset compared to age-matched control BALB/c. Lymphocytes from the hyporesponsive mice specifically suppressed the proliferative responses of control BALB/c to DBA/2 but not to AKR. The data indicate that maternal hyperimmunization can induce tolerance by a mechanism involving intrathymic selection of suppressor cells which can be combined with a negative selection of helper cells.     

Antibody Response in C3H Mice Injected with CBA Lymphoid Cells As Detected by Membrane Immunofluorescent Staining

Lymphocytes from CBA mice are strongly responsive to cells from the H-2 identical strain C3H in vitro (MLC), whereas C3H lymphocytes are poorly reactive against CBA cells. Immunization of CBA mice with C3H lymphocytes did not yield any detectable specific antibodies. On the other hand, C3H mice immunised with CBA cells produced specific antibodies as detected by membrane immunofluorescence. Mice of the strains AKR and DBA/2 also possess this specific membrane alloantigen. We thus conclude that it is now possible to discriminate among lymphocytes of CBA and C3H origin.     

Protection of newborn mice from graft versus host disease by maternal pre-immunization

Alloimmunization of BALB/c (H-2d) female mice with allogeneic spleen cells from C57BL/6 (H-2b) or CBA/H (H-2k) mice protects BALB/c offspring from graft-versus-host disease (GVH-D) following neonatal intraperitoneal inoculation of high doses of spleen cells respectively of C57BL/6 or CBA/H strains of mice. The mice survived GVH-D over one year after the allogeneic inoculum 24-48 h after birth and they did not show any signs of GVH reaction nor splenomegaly. We show that this phenomenon is antibody mediated and affects the developing immune system of the foetus. Repeated immunization of virgin female BALB/c with anti-H-2b or anti-H-2k antisera (Ab1) can equally abrogate GVH-D in their newborn offspring challenged at 24-48 h after birth with allogeneic spleen cells of H-2b or H-2k phenotype. Our results demonstrate that protection from GVH-D is not specific to the immunizing strain and occurs when the neonatal mice are challenged with C57BL/6 or CBA/H spleen cells. There is thus crossreactivity of tolerance against H-2 specificities. In this study we also report on the in vitro cellular immune responses of the surviving GVH-resistant mice and demonstrate that these responses against both the challenge and third party lymphocytes are impaired.     

H-2-linked murine cytotoxic T cell responses specific for sendai virus-infected cells.

CBA (H-2k) mouse-derived lymphochoriomeningitis virus and herpes simplex virus-specific cytotoxic T lymphocytes lyse virus-infected target cells compatible on either the H-2k or H-2D region. In contrast, CBA, C3H and AKR (H-2k) mouse-derived sendai virus-specific cytotoxic T lymphocytes (CTL) fail to lyse H-2D-compatible virus-infected cells. A similar lack of H-2D region-associated lytic activity was found with C57BL/6 and C57BL/10 (H-2b) mice as well as with the recombinants B10.A (2R) [Kb-Db] and B10.A (4R) [Kk-Db]. On the other hand, BALB/c (H-2d) mice and A/J (H-2a) mice do generate H-2Dd-associated sendai virus-specific CTL. These results are in contrast to those obtained with (CBA X BALB/c)F1 and B10.HTT [Ks-Dd] mice, which failed to mount Dd region-associated CTL responses. It is concluded that D region-associated sendai virus-specific CTL responsiveness varies with the H-2 genotype of the responder cells.     

Three-dimensional collagen matrices as cell culture substrata affect the generation of alloreactive cytotoxic T lymphocytes

Primary one-way mixed lymphocyte cultures (MLC) of C3H/He responder and DBA/2 stimulator were performed in three-dimensional (3-D) collagen matrices and the generation of alloreactive cytotoxic T cell (CTL) responses was compared to those in MLC which were done on usual plastic surfaces or on collagen-coated plastic surfaces. MLC in the 3-D collagen matrices were found to generate strong CTL responses. Flow cytometric analysis of Lyt-2 and L3T4 antigen expressions on the effector cells showed that the Lyt-2/L3T4 ratios were substantially higher in the 3-D collagen matrices, and that a larger proportion of the cells in the 3-D collagen matrices were Lyt-2+ lymphoblasts. These results indicate that the milieu of the 3-D collagen matrices favors the proliferation of Lyt-2+ lymphocytes, and suggests that cell-to-matrix interactions in 3-D collagen matrices may play a regulatory role in the maturation process of alloreactive CTLs.     

Immune response gene regulation of the humoral immune response to Porphyromonas gingivalis fimbriae in mice.

Among various strains of mice immunized orally with Porphyromonas gingivalis fimbriae and adjuvant GM-53 in liposomes, BALB/c and DBA/2 mice (H-2d) were found to be high responders to the fimbriae, CBA/J and C3H mice (H-2k) were intermediate, while C57BL/6 mice were low responders in terms of serum IgG and salivary IgA responses. Furthermore, humoral immune responses were examined using congeneic mice of B10 background showing different H-2 haplotypes, and it was revealed that B10.D2 mice (H-2d), followed by B10.BR (H-2k), responded well to antigenic stimulation of the fimbriae, while C57BL/10 mice (B10, H-2b) were low responders to the fimbriae. Hybrids between BALB/c and C57BL/6 mice were found to reflect a phenotype of low responders. Thus, the humoral immune responses to P. gingivalis in mice are restricted by H-2 haplotype.     

Cell-mediated immunity in experimental murine dermatophytosis. I. Temporal aspects of T-suppressor activity caused by Trichophyton quinckeanum.

Cutaneous dermatophyte infections caused by Trichophyton quinckeanum were established in various strains of mice. All congenic BALB/B (H-2b), BALB/c (H-2d) and BALB/K (H-2k) strains showed high susceptibility to dermatophyte infection. Susceptibility is independent of MHC phenotype, since other strains with corresponding H-2 haplotypes such as C57BL/6 (H-2b), DBA/2 (H-2d) and CBA (H-2k) were resistant to the disease. During the acute phase of infection in BALB/c mice, the in vitro blastogenic responses of regional lymph node cells was suppressed. Suppression was observed for both the T cell mitogens concanavalin A and phytohaemagglutinin, and for the B cell mitogen lipopolysaccharide. A series of cell-mixing experiments revealed that lymph node cells from infected mice were able to suppress the T cell and B cell mitogenic responses of lymph node cells from normal mice. Suppression was mediated by T cells and abrogated by treatment of lymph node cells with either monoclonal anti-thy-1.2 or anti-Ly-2.2 and complement. In this report, we discuss the possibility that T-suppressor mechanism mediated by cells bearing the Ly-2+ phenotype and activated during dermatophyte infection may determine the course of the disease.     

Weak Graft-Versus-Host Response of CBA Lymphocytes Against the H-2-Identical Strain C3H

Lymphocytes from CBA and C3H mice were tested for graft-versus-host (GVH) reactivity in C3H × CBA recipients. Three different GVH assays were used, namely, the spleen enlargement test, the popliteal lymph node assay, and inhibition of allogeneic bone marrow proliferation CBA lymphocytes showed only weak reactivity against C3H X CBA tissues in all these tests, while there was strong reactivity in H-2-different F₁ hybrids. The capacity of C3H and CBA cells in producing a host-versus-graft (HVG) reaction was also tested. It was observed that CBA cells injected into the foot pads of C₃H mice induced an enlargement of the local popliteal lymph node that was as great as when C3H cells were injected into CBA mice. On the other hand, cells from the H-2-different strain 57B1 induced a considerably smaller HVG reaition when injected into CBA mice. It is concluded that antigens that strongly stimulate mixed lymphocyte cultures do not always stimulate allogeneic lymphocytes to strong GVH reactions in vivo.     

Maturation of the filaria Litomosoides sigmodontis in BALB/c mice; comparative susceptibility of nine other inbred strains.

When inoculated subcutaneously, the infective larvae of L. sigmodontis undergo complete development and produce a patent microfilaraemia in mice of the BALB background (BALB/c, BALB/K and BALB/B, with respectively the H-2d, H-2k et H-2b haplotypes). The most susceptible strain is BALB/c with all mice harbouring adult filariae and 47% of mice presenting with a patent microfilaraemia. Mice with the B10 background (B10, B10Br and B10D2, with respectively the H-2b, H-2k et H-2d haplotypes) are almost completely resistant to infection. Adult filariae were recovered from all mice of the CBA/Ca, CBA/HN, C3H/HeN, DBA/2N strains. However, the site and structural development of the parasite varied in each strain. Absence of microfilaraemia is associated with absent or abnormal spicules, reduced number of female filariae and small size of female filariae. These results show that the Major Histocompatibility Complex only modulates the developmental pattern of filariae within the limits imposed by background genes. Male CBA/HN and C3H/HeN were more susceptible to infection than female mice. Inverse phenomenon was observed with strains BALB/c; and, no host sex effect was seen in DBA/D2N.     

过继转输胚胎抗原耐受T细胞诱导自然流产孕鼠母胎免疫耐受

探讨过继转输胚胎抗原耐受T细胞对小鼠自然流产模型妊娠预后及宿主孕鼠免疫细胞对父系抗原免疫耐受状态的影响。以CBA/J×BALB/c为正常妊娠模型 ,CBA/J×DBA/ 2为自然流产模型 ,将自然流产模型CBA/J孕鼠于孕 4d (着床期 )分别腹腔注射大鼠抗小鼠CD80和CD86mAb或大鼠同型IgG。于孕 9d ,应用免疫磁珠阴性分选三组孕鼠的脾脏T细胞 ,并将三组T细胞分别转输至孕 4d的CBA/J×DBA/ 2孕鼠。于宿主孕鼠孕第 9天 ,采用单向混合淋巴细胞反应分析宿主孕鼠脾脏免疫细胞对父系抗原的增殖能力 ,并用流式细胞术分析经父系抗原刺激的宿主孕鼠脾脏T细胞内IL 2表达水平。于孕 1 4d分别观察宿主孕鼠的胚胎吸收率。结果显示 ,过继转输胚胎抗原耐受T细胞和转输正常妊娠模型孕鼠的T细胞均可诱导宿主孕鼠脾脏免疫细胞对父系抗原的增殖能力及IL 2的表达显著下降 (P <0 0 5 ) ,孕 1 4d胚胎吸收率也显著下降 (P <0 0 5 )。这些结果表明 ,于孕早期过继转输胚胎抗原耐受T细胞和转输正常妊娠模型孕鼠的T细胞能诱导宿主孕鼠母 胎免疫耐受 ,防止母体对胚胎的免疫排斥 ,从而使自然流产模型的妊娠预后达到正常妊娠水平。     

Reduced Capacity to Produce Specific 'Effector' Cells after Injection of CBA Mice with C3H Cells

Lymphocytes from mice of strain CBA are strongly MLC-responsive to lymphocytes from the H-2 compatible- but M-antigen-incompatible strain C3H. This strong reactivity disappears after infusion of CBA mice with C3H lymphocytes. This study shows that the host-versus-graft reactivity (swelling of local lymph node after antigen injection) is specifically reduced after injection of CBA mice with C3H × CBA spleen cells However, lymphocytes from such mice showed a specifically increased GVH reactivity (inhibition of erythroid cell growth) compared with lymphocytes from unimmunized mice. Lymphocytes from normal CBA mice showed a high proliferative rate in the spleens of irradiated C3H × CBA mice. Such 'educated' cells showed strongly increased specific GVH reactivity. Lymphocytes from CBA mice previously injected with C3H×CBA cells showed reduced capacity to proliferate when injected into irradiated C3H × CBA hybrids and a poor capacity to develop new 'effector' cells reactive against C3H × CBA bone marrow target cells. The results indicate that the presence of specifically 'MLC responsive' lymphocytes in a lymphoid cell population is a prerequisite of its production of 'effector' cells able to respond in this GVH assay     

The glycosphingolipid globoside as a serological marker on cytolytic T lymphocyte precursors and alloantigen-responsive proliferating T lymphocytes in murine spleen

Biochemical analyses of murine lymphocytes have shown that the glycosphingolipid globoside (Glo) is present exclusively on alloantigen-stimulated murine T lymphocytes (Gruner, K. R., Van Eijk, R. V. W. and Mühlradt, P. F., Biochemistry 1981. 20: 4518). An anti-Glo antibody has now been raised in rabbits immunized with purified antigen. Most activity was recovered in the IgM fraction. The specificity of the antibody was ascertained in an enzyme-linked immunosorbent assay with purified glycosphingolipids bound to the solid phase. In antibody-dependent complement lysis experiments the anti-Glo eliminated about 20% of nylon wool-nonadherent splenic T cells of CBA/J mice. To determine the functional identity of these Glo+ cells, the effects of Glo+ cell elimination on mitogen stimulation with concanavalin A and lipopolysaccharide, as well as the effects on the mixed lymphocyte culture (MLC) reaction and cell-mediated lympholysis with mitomycin-treated DBA/2 splenocytes as stimulator cells were studied. Whereas lipopolysaccharide stimulation was not affected by elimination of Glo+ cells, there was a slight inhibitory effect on the concanavalin A stimulation, and a severe inhibition of the MLC reaction and the generation of H-2d-specific cytolytic T lymphocytes. Addition of interleukin 2 increased the MLC reaction, but interleukin 2-saturated cultures were also severely inhibited by anti-Glo and complement treatment. Combined treatment with anti-Glo and anti-Lyt-1 or anti-Lyt-2 antibodies, and determination of cytolytic T lymphocyte precursor frequencies in limiting dilution cultures after Glo+ cell elimination showed that a large proportion of T cells proliferating in a primary MLC are Lyt-1+,2+,3+Glo+, whereas in secondary MLC they are Lyt-1+,2-,3-,Glo+. Fifty % of the cytolytic T lymphocyte precursors in primary as well as secondary MLC are Glo+. The Glo marker is lost upon differentiation to cytolytic T lymphocyte effector cells. It is discussed herein that Glo is a marker for alloantigen-stimulated precursor T lymphocytes of both helper and cytolytic T cells.     

Influence of the genetic background and parasite load of mice on the immune response developed against nymphs of Ixodes ricinus

The immune response of BALB/c (H-2d), DBA (H-2d), C57BL/6 (H-2b), C3H (H-2k), CBA (H-2k), SJL (H-2s), and FVB (H-2q) mice infested once with 15 nymphs of Ixodes ricinus is polarized toward Th2 as suggested by cytokines produced by lymph node cells stimulated with concanavalin A. The parasite load does not influence the polarization of the immune response as observed in BALB/c mice, which developed a Th2 response when infested with 5 or 45 nymphs. As assessed by attachment and weights of engorged nymphs, no resistance was acquired by BALB/c, C57BL/6, or C3H mice undergoing three successive infestations. However, these mice produced a gradual increase in IgE. Received: 8 December 1998 / Accepted: 22 December 1998     

Immunity Parameters in Mice of Different Strains

Quantitative composition and functional activity of immunocompetent cells differ in mice of different strains. The counts of T cells in the bone marrow and spleen, proliferative activity of T cells in the spleen, levels of IL-2 and IL-10 production by splenic T cells, number of antigen-specific T cells and their functional activity are low in C57Bl/6, BALB/c, and CC57W mice and high in CBA/CaLac, DBA/2, and C3H animals. Low phagocytic activity of peritoneal macrophages was detected in BALB/c and CC57W mice and high activity in C3H animals. The content of antibody-producing cells in the spleens of C57Bl/6, BALB/c, and CC57W mice is higher than in CBA/CaLac, DBA/2, C3H, A/SN, and AKR/JY mice. Functional activity of B cells is lower in BALB/c and CC57W compared to CBA/CaLac and DBA/2 mice. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 140, No. 8, pp. 189–191, August, 2005     

Ly-49D transfected NK cells show reduced interleukin-10 production in response to H-2d and increased lytic activity: implication by interaction using class I MHC alloantigen+ target cells in pregnancy

PROBLEM: Mating of CBA/J (H-2k) with DBA/2 (H-2d) males leads to a high rate of spontaneous resorption (about 40%), which is not seen in other mating combinations, such as CBA/J X BALB/c. The activation of natural killer cells (NK cells) seems to be a key mechanism for the maternal-fetal intolerance in allogeneic pregnancy, and recurrent spontaneous abortion. The effect of expression of the NK cell activating receptor Ly49D recognizing BALB/c or DBA/2 class I MHC was investigated. METHOD OF STUDY: Intracellular interleukin (IL)-100 production was detected and target cell survival rates were calculated after 22 hr coincubation of rat NK cells transfected, or not. with a murine Ly49D receptor, with either male BALB/c or male DBA/2 splenocytes, by using flow cytometry. RESULTS: Ly49D negative rat NK cells produced 13.7% more IL-10 than Ly49D positive rat NK cells, and more splenocytes were killed by Ly49D transfected rat NK cells (survival rate 2.45%) than by Ly49D negative rat NK cells (survival rate 4.36%). CONCLUSION: After physiological stimulation with BALB/c or DBA/2 splenocytes, rat NK cells are able to synthesize IL-10. Recognition of mouse splenocyte major histocompatibility complex (MHC) by Ly49D mice receptor decreased IL-10 production. The observed increase in killing activity might be a result of this phenomenon. NK cell activation via the Ly49D receptor might play an important role in pregnancy failure, but cannot explain why CBA/J X DBA/2 matings are abortion prone, and CBA/J X BALB/c matings are abortion resistant.     

Immune reactions in different mouse strains

The responses of immunocompetent cells to thymus-dependent antigen differ in mice of different strains. Immunization stimulated phagocytic activity of peritoneal macrophages in CBA/CaLac, DBA/2, and BALB/c mice and suppressed it in CC57W mice. By the formation of antibody-producing cells in the spleen in response to thymus-dependent antigen DBA/2 and CBA/CaLac mice can be classified as high responders, BALB/c mice as medium-responders, and C57Bl/6 and CC57W mice as low responders.     

Reduced stathmin-1 expression in natural killer cells associated with spontaneous abortion

Female CBA/J mice impregnated by male DBA/2J mice (CBA/J×DBA/2J matings) are prone to spontaneous abortion, although the reason for this is unclear. In this study, the stathmin-1 expression pattern was evaluated in uterine natural killer (uNK) cells purified from CBA/J×DBA/2J matings. Results were compared with those in a CBA/J×BALB/c control group that yields successful pregnancies. The mean ± SD percentage of stathmin-1(+) cells in the CD49b(+) uNK cell population was lower in CBA/J×DBA/2J mice (0.7% ± 0.4%) than in control CBA/J×BALB/c mice (4.9% ± 1.5%, P < 0.01) using flow cytometry, and the intracellular stathmin-1 level in uNK cells was lower in CBA/J×DBA/2J mice than in control mice using Western blot analysis. Co-localization of lectin from Dolichos biflorus agglutinin (DBA-lectin) and stathmin-1 was confirmed using multivision immunohistochemical analysis. The frequency of stathmin-1(+)DBA-lectin(+) cells was lower in CBA/J×DBA/2J mice than in CBA/J×BALB/c mice. A similar trend in the frequency of stathmin-1(+)CD56(+) cells was seen in patients with unexplained spontaneous abortion compared with normal early pregnancy. A neutralizing antibody against stathmin-1 further increased the percentage of embryo loss in CBA/J×DBA/2J matings. These results provide evidence that stathmin-1 expression in uNK cells at the maternal-fetal interface may help modulate uNK cell function and may be beneficial for a successful pregnancy.     

Clones of cytotoxic lymphocytes in culture: the difference in specificity between stimulated and nonstimulated cytotoxic lymphocytes.

The specificity of individual clones of cytotoxic lymphocytes (CL) which develop upon stimulation by semi-allogeneic cells, have been examined and compared with the specificity of CL clones which develop spontaneously when normal spleen cells are cultured without stimulator cells. It was found that the specificity of stimulated clones was different from the specificity of 'spontaneous' clones. Specifically stimulated clones from CBA cells cultured with (CBA x DBA)F1 stimulator cells, did not discriminate between the H2dP815 and H2dDBA splenic blast, and lysed both targets. In contrast, spontaneous CL clones from cultures of CBA or (CBA x DBA)F1 spleen cells were found to lyse either P815 or DBA blast, but not both. The results indicated that the spontaneous CL clones were not a representative sample of the the total pool of CL.     

Streptococcal group A carbohydrate antibodies in mice: evidence for strain differences in magnitude and restriction of the response, and for thymus dependence

Immunization with streptococcal group A vaccines revealed significantly higher immune responses in BALB/c than C57BL/6, CBA, and DBA/2 mice. In general, the antibody response of BALB/c mice not only was markedly higher than that of the other strains, but also showed a high degree of restriction. With two exceptions, all immune sera of the BALB/c mice exhibited bands in very similar electrophoretic positions. On the other hand, the three other strains were low responders which, except for the DBA/2 mice, rarely demonstrated clear-cut bands in the immune sera. Studies with thymusless (nude) mice carrying 50 – 85% BALB/c genome identified the streptococcal group A polysaccharide, presented on whole bacteria, as a thymus-dependent antigen. These nude mice, after reconstitution with thymus cells, produced group A antibodies restricted to two antibody bands extremely similar to those of BALB/c mice. High levels of antibody were produced in mice reconstituted with low responder DBA/2 thymus cells, indicating that the magnitude of the response is entirely controlled by mechanisms at the bone marrow cell level. Magnitude and restriction of the antibody response to this antigen are not determined by the H-2 allele and are not related to the Ig^a allotypic marker on the heavy chain.     

Immunogenetic studies of spontaneous abortion in mice. III. Non-H-2 antigens and gestation

CBA/J (H-2k) females, mated with DBA/2 J (H-2d) or DBA/1 J (H-2q) males, exhibit a high rate of fetal resorption. In contrast, when H-2 identical CBA substrains (i.e. CBA/Ca and CBA/N) are used, this phenomenon is not observed. On the other hand, before mating with DBA/2 J males, pre-immunization of CBA/J females with spleen cells coming from BALB/c J or (DBA/2 x BALB/c J) F1 males (and not from other H-2d identical males whatever their Mls alleles) has significantly decreased the fetal resorption rate. Thus, immunization against determinants other than classical H-2d (K, I, D, L) antigens (transmitted as a dominant character and different from Mls determinants) can elicit anti-abortive effects. Furthermore, it was observed that the spleen cell endowed with the anti-abortive effects was neither a T nor a B lymphocyte. In contrast, peritoneal cells were able to reproduce the phenomenon, indicating that it may be mediated by a cell of the macrophage-monocyte lineage. Finally, a first gestation was substituted for allo-immunization of CBA/J females. The anti-abortive effects of a first pregnancy by BALB/c J males (and not by other H-2k syngeneic or H-2d allogeneic males) was observed in the course of a second pregnancy sired by DBA/2 J males. These data can be interpreted in terms of maternal recognition of an antigen present on both macrophages and trophoblast cells and necessary for a successful gestation, which is coded for by genes outside the K, I, D, L regions.