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Biomedical subjects

A Coutinho

Publications and source records attributed to A Coutinho.

At least 181 records · Page 10Linked to original sources
PubMed1988

Long-lasting thymus-independent immune responses to anti-idiotype lipopolysaccharide conjugates require continuous B cell renewal.

In the present investigation, we have studied the antibody response of both C57BL/6 lipopolysaccharide-responder mice (B6 LPS-R) and C57BL/10ScCr LPS-nonresponder mice (B10.Cr LPS-NR) upon immunization with copolymers of LPS and either Ac38 or Ac46 anti-idiotypes. As both strains of mice studied are of the Ighb allotype the antibody response obtained was quantified by estimating the serum levels of the complementary idiotype, B1-8, in immunized mice. The results show that while the injection of the LPS-R strain B6 with copolymers of LPS anti-idiotype resulted in a long-lasting idiotype response the immunization of the LPS-NR strain, B10.Cr, resulted in virtually no response. This finding allowed us to perform cell transfer experiments to study the response of isolated cells from B6 mice transferred into LPS-NR hosts. Using such a cell transfer model, the results obtained suggest that long-lasting idiotype responses observed in the LPS-R strain require continuous B cell production in bone marrow. The relevance of this finding to the understanding of the long-lasting (fluctuating) antibody responses to nondegradable thymus-independent polysaccharides is discussed.

Animals↗
PubMed1988

T cell dependence of the "natural" autoreactive B cell activation in the spleen of normal mice.

A high frequency of clonal precursor B cells producing lytic antibodies to syngeneic erythrocytes treated with bromelain (BMRC) is revealed in normal mouse spleen cells by lipopolysaccharide-driven limiting dilution analysis. All such specificities are recovered as activated blasts after density gradient fractionation, the "small lymphocyte" pool being depleted of anti-BMRC reactivities. In contrast, the spleens of athymic (nude) mice contain undetectably low frequencies of these specificities in either lymphocyte compartment. Transfer of relatively low numbers of normal syngeneic splenic T lymphocytes to adult nude mice restores the high frequency of anti-BMRC clonal precursor B cells, again in the activated, but not in the resting spleen cell fractions. Large total T cells are more than tenfold better than resting T cells in reconstitution potential, as are enriched CD4+ as compared to CD8+ cells which are practically devoid of activity in this respect. These results apply exclusively to B cells at a differentiative stage that allows for extensive clonal expansion, since there is a marked difference between the frequency of clonal precursors determined by limiting dilution analysis and the frequency of Ig-secreting plaque-forming cells of the same specificity, and induced by the same mitogen in short-term cultures. The implications of these findings for the physiology of autoreactivity and repertoire selection in the compartment of perinatal B cells are discussed.

Animals↗
PubMed1988

Back-stimulation of B lymphocytes binding to helper T cell surface antigens.

Helper T cells directed to B cell surface determinants activate their "targets" into polyclonal antibody production in vitro and in vivo, but B cells which bind to epitopes on the helper cell surface are preferentially induced. Furthermore, "anti-helper" B cell activation also occurs by "back-stimulation", that is even when the responding B lymphocytes are not specific targets for the inducing helper cells, as long as these are simultaneously activated by appropriate interactions with presenting cells. In our experimental systems, this is the only condition where "bystander" activation can be recorded. These findings suggest mechanisms of auto-antibody production associated with unrelated helper cell activity in vivo.

Animals↗
PubMed1988

A phenotypic and functional analysis of long-lived B and T lymphocytes.

The lymphocyte composition of spleen, lymph nodes, bone marrow, and thymus of mice submitted to hydroxyurea treatments for four consecutive days was studied. The treatment selects for small lymphocyte populations that represent between 4 and 20% of control numbers in the various organs. Spleen and bone marrow contain the same B cell population with a low IgM, high IgD, low I-E phenotype, which respond to LPS at control clonal frequencies. The T cell compartment is equally depleted, and the lymphocytes remaining contain frequencies of clonable cells in response to mitogens and IL-2 that are comparable to those detected in normal spleen cells. Overall, the results suggest that only a minor fraction of all lymphocytes in a normal young adult mouse have life spans longer than 4 days.

Animals↗
PubMed1988

Most B cells in acute Trypanosoma cruzi infection lack parasite specificity.

The specificity of B lymphocytes activated in the acute phase of murine Trypanosoma cruzi infection was analysed in a panel of immunoglobulin-secreting hybridomas derived by fusion of lymph node cells 7 days after intraperitoneal parasite inoculation. The immunoglobulin isotype distribution of the hybrids reflected the total plaque-forming cell (PFC) response in the animal at this point, with a predominance of IgG2a, IgM, and IgG2b. Screening of the hybridoma antibodies on parasite antigens by three independent methods (western blot, ELISA, and immunofluorescence) revealed only one (out of a total of 51) that bound a parasite molecule with an apparent molecular mass of 180 kDa. In contrast, antibodies of both IgM and IgG classes were found to react with a panel of autologous antigens. These results establish that most B cells activated by T. cruzi infection are not specific for parasite antigens and therefore indicate the relevance of analysing the totality of host responses to infection.

Acute Disease↗
PubMed1988

On the validity of using lipopolysaccharide-driven limiting dilution systems for clonable B-cells to analyse functional antibody repertoires.

Analyses of B-cell repertoires by mitogen-driven limiting dilution systems (LDA) followed by specific antibody detection on immobilized antigens (ELISA), while constituting the only method available to determine absolute frequencies of any given specificity, provide no indications as to the functional ability of clonal precursors scored as positive to actually respond to antigen in vivo. We have now addressed this question by quantitating dextran alpha 1----6-specific clonal precursors among small, resting, and large activated B cells in the spleens of mice, before and after priming with optimal doses of antigen. The results demonstrate that virtually all B cells scored as antigen-specific in LDA/ELISA systems do respond to antigen and undergo blast transformation after priming in vivo.

Animals↗
PubMed1988

The V beta 8 gene family is preferentially used by 'naturally' activated T cells.

Receptors encoded by the V beta 8 gene family detected by the monoclonal antibody F23.1 are expressed among 'naturally' activated T cells in normal spleen at frequencies significantly higher than in the total CD4+ and CD8+ cell populations. The positive selection of these clones into 'natural' T-cell activity could be the reason for the high frequencies of cells expressing V beta 8 genes. This phenotype is strain-dependent.

Animals↗
PubMed1988

Membrane expression of IgG but not maturation to secretion requires DNA replication.

In this work we have addressed two questions. Does switch recombination occur before membrane expression or only concomitantly with induction to high rate synthesis and secretion of IgG? Does interleukin-4 induce switch to IgG1 or maturation of already switched cells? To answer these questions we used the DNA synthesis inhibitor hydroxyurea (HU) to analyze the requirements for DNA replication in the induction of membrane expression or high rate secretion of all IgG subclasses by cultures of mouse spleen cells stimulated by lipopolysaccharide (LPS) and supplemented with IL-4, the absolute numbers of cells bearing membrane bound IgG was always decreased by HU, indicating that immunoglobulin class switching requires DNA replication. IL-4 did not augment the numbers of cells expressing any IgG isotype. In contrast, the number of high rate secretors of all IgG isotypes was not affected by HU, except in the case of IgG3 and IgG2b shortly after stimulation. Addition of IL-4 resulted in an increased number of secretory cells, and also this effect was resistant to HU. Thus, for any isotype, treatment with HU or IL-4 increased the frequency of secretory cells relative to the surface positive population. This data indicates that: 1) IL-4 is a broad range, non isotype-specific maturation factor for LPS-activated B cells; 2) induction to high rate secretion has a negative effect on proliferation; and 3) immunoglobulin class switch, but not induction to secretion of any immunoglobulin isotype, requires DNA replication, suggesting that switch recombination had to occur before expression of IgG in the membrane-bound form.

Animals↗
PubMed1987

Suppression of polyclonal antibody production in Trypanosoma cruzi-infected mice by treatment with anti-L3T4 antibodies.

Acute Trypanosoma cruzi infection of mice results in a very marked polyclonal activation of B and T lymphocytes, accompanied by high numbers of Ig-secreting PFC and lectin-dependent effector CTL. Treatment of mice with monoclonal anti-L3T4 antibodies from the time of infection (days 0, 4, and 8) completely suppresses the polyclonal PFC response and CTL generation. Treatment of nude mice with antibody does not alter the lipopolysaccharide-induced polyclonal PFC response, and it only modulates the isotypic profile of the PFC response to T. cruzi infection, without reducing its magnitude. Furthermore, antibody-treated, T. cruzi-infected euthymic mice do not develop the typical B cell blastogenic response, but show high numbers of activated Lyt-2+ lymphoblasts in the spleen. These results indicate that effector cell generation in T. cruzi-infected mice is predominantly helper T cell-dependent.

Animals↗
PubMed1987

The immune response to bacterial dextrans. V. A "dominant" idiotype in IgCHb mice.

A battery of syngeneic monoclonal anti-idiotypic antibodies was prepared against a monoclonal C57BL/6 anti-Dextran B512 antibody (17-9). Two such anti-idiotypes were shown to bind to sites on the 17-9 molecule which are related to the Dex-binding site and were used to characterize the anti-Dex antibody response in a number of inbred mouse strains. The results show that the 17-9 idiotype is recurrently expressed by all mice carrying the IgCHb haplotype, regardless of H-2 or background genes, and that this idiotype accounts for roughly one-half of the primary, specific response to Dex B512 in C57BL/6 mice. Backcross analysis confirmed the allotype-linkage of idiotype expression in the antibody response. Mice carrying other allotypes, however, had detectable levels of the 17-9 idiotype in normal sera, which was not associated with anti-Dex antibody activity and was not raised by specific immunization. Together with previous observations, these results characterize a second "recurrent" idiotype in the anti-alpha,1-6 response of IgCHb mice, both of which are expressed in the normal serum of all mouse strains tested.

Animals↗
PubMed1987

Long-lived B cells: mitogen reactivity as a tool for studying their life-spans.

A population of noncycling small lymphocytes has been selectively enriched in mice by treatment with hydroxyurea (HU) over 4 consecutive days. This cell population, present in the spleen of HU-treated mice, contained a high frequency of lipopolysaccharide (LPS)-reactive cells and this could be used as a functional marker to follow their persistence after transfer to LPS-nonresponder recipient mice. The results reported here show that cells selected after a long-term HU treatment have considerably less decay, after transfer in recipient mice, than normal spleen cells. Furthermore, they demonstrate that HU-selected B cells survive in the recipient mice for periods up to 3 months, and are, consequently, true long-lived lymphocytes.

Animals↗
PubMed1987

Isotypic pattern of the polyclonal B cell response during primary infection by Plasmodium chabaudi and in immune-protected mice.

The primary infection by P. chabaudi induces an increase of the numbers of splenic immunoglobulin (Ig)-secreting B cells in both athymic and euthymic BALB/c mice. The isotypic pattern of the polyclonal response is restricted only in euthymic mice where IgG2a, IgG2b and IgM plaque-forming cells (PFC) predominate. In immunized animals, protected against a parasite challenge, the isotypic pattern of splenic PFC is completely different, the IgG1 and IgM isotypes constituting the main part of the response. Reinoculation of immune-protected animals induces a PFC response which is dose dependent and accentuates the characteristic isotypic profile of the immune-protected mice.

Animals↗