Electrophoresis of buffalo (bos bubalis) serum proteins including immunoglobulins.

Antigenic components of buffalo (Bos bubalis) serum, which were also components of buffalo colostrum, seminal plasma, milk whey, saliva, and tears, were investigated by the ager gel diffusion test and immunoelectrophoresis. Immunoglobulins of buffalo serum were identified by immunoelectrophoresis employing rabbit-anti-buffalo serum and rabbit-anti-buffalo gamma-globulin. Based on immunoelectrophoretic patterns immunoglobulin G (IgG), IgGA, and IgM were detected both in the serum and colostrum of buffaloes. Tears contained both IgG and IgM. Cross-reactions of buffalo serum with seminal plasma, saliva, and milk whey were observed only in the IgG region. By polyacrylamide gel electrophoresis, lipoprotein (5.2% +/- 0.41), IgM (11.4% +/- 3.1), IgG (9.4% +/- 0.98), haptoglobin 21.8% +/- 3.73), transferrin (10.4% +/- 2.15), ceruloplasmin (7.8% +/- 1.3), postalbumin (20.8% +/- 2.09), and albumin (13.7% +/- 0.75) were identified provisionally.     

Use of intravenous polyclonal immunoglobulins in autoimmune and inflammatory disorders]

Initially used for the treatment of immunodeficiencies, intravenous immunoglobulins (IVIg) have increasingly been used as immunomodulatory agent in autoimmune and inflammatory disorders. The mode of action of IVIg is enigmatic, probably involving Fc-dependent and/or F(ab')2-dependent non-exclusive mechanisms of action. IVIg broadly interacts with the different components of the immune system: cytokines, complement, Fc receptors and several cell surface immunocompetent molecules. IVIg also has an impact on effector functions of immune cells. These mechanisms of action of IVIg reflect the importance of natural antibodies in the maintenance of immune homeostasis. We discuss here the recent advances in the understanding of immunoregulatory effects of IVIg, and we pointed out the need of new strategies to overcome the predicted increasing worldwide shortage of IVIg.     

Suppression of experimental autoimmune encephalomyelitis by intravenously administered polyclonal immunoglobulins

Experimental autoimmune encephalomyelitis (EAE) was induced in Lewis rats either by active immunization with myelin basic protein (MBP) or by adoptive transfer using anti-MBP specific CD4(+)T cells. Treatment with human polyclonal immunoglobulins (IgG) effectively suppressed active EAE. Time-dependent experiments demonstrated that the effect of IgG was manifested only when treatment was given immediately after immunization; administration from day 7 after disease induction did not suppress the disease. In the adoptive transfer model of EAE, IgG had no effect in vivo. However, pretreatment in vitro of the antigen-specific T-cells with IgG inhibited their ability to mediate adoptive EAE, as it did in active EAE. Similarly, in vitro IgG pretreatment of the antigen-specific T-cells suppressed the proliferative response to MBP. Fluorescent Activated Cell Sorter (FACS) analysis demonstrated the binding of IgG to activated T-cell lines that was inhibited by soluble Fc molecules. The differential effects of IgG on active EAE and on the adoptive transfer of EAE suggest that IgG in vivo can suppress disease by acting during the early phase of the immune response which involves naive T cells. The inhibition of T-cell proliferation and adoptive transfer of EAE by incubation of T cells in vitro appears to require higher concentrations of IgG than those obtained in vivo.     

Induction of oral tolerance in experimental antiphospholipid syndrome by feeding with polyclonal immunoglobulins

Intravenous immunoglobulins (IVIG) contain a wide spectrum of anti-idiotypes associated with autoimmune diseases. Since part of these anti-idiotypes may bear an internal image of the eliciting antigen, IVIG might be suitable for induction of oral tolerance. In the current study we attempted to induce tolerance in an experimental model of anti-phospholipid syndrome (APS) by oral administration of IVIG. Naive mice were fed with IVIG, or anti-beta 2GPI-specific anti-idiotypic IVIG(alpha Id). Significantly diminished humoral response was noted in mice IVIG/ IVIG-F(ab')(2)or IVIG(alpha Id)-tolerized mice, accompanied by a significant attenuation of clinical manifestations. The maximal effect was achieved in the mice tolerized before disease induction. Abrogation of T lymphocyte proliferation to beta 2GPI was detected in the mice fed with IVIG prior to beta 2GPI immunization, mediated by TGFbeta and IL-10 secretion. The tolerance induced by IVIG-feeding was nonspecific and could be adoptively transferred to syngeneic mice by CD8alpha (+) cells. These CD8alpha (+) T cells, were found to secrete high levels of TGFbeta and IL-10. In summary, IVIG-induced oral tolerance has a nonspecific immunomodulatory effect in experimental APS, mediated by TGFbeta and IL-10-secreting CD8alpha (+) cells. Our results point to a possible application of IVIG in the induction of oral tolerance against various autoimmune diseases.     

免疫透射比浊法测定血清免疫球蛋白

本文报道免疫透射比浊法测定血清免疫球蛋白,批内CV小于2.0％,批间CV小于3.5％,表明该法简便,快速,精密度和准确度高,重复性好。     

Human secretory immunoglobulins. II. Salivary secretions from individuals with selectively excessive or defective synthesis of serum immunoglobulins

Increased amounts of IgG were transmitted into the salivary secretions of patients with elevated serum levels of this immunoglobulin. Both its glandular and extra-glandular transfer apparently depended upon passive diffusion or epithelial `leakage'.

High serum levels of IgM (in macroglobulinaemia), and especially local synthesis of this immunoglobulin (in IgA deficiency), enhanced its transfer into the saliva. The transmission through secretory epithelium (glandular transfer) seemed to be an active or selective process, probably dependent upon specific `transfer sites' in the heavy polypetide chains of IgM.

The transmitted immunoglobulin (secretory IgM) was physicochemically and immunochemically similar to its 19S counterpart in serum. In IgA-deficient secretions no significant association between immunoglobulin components and secretory piece (SP) could be detected; the latter was physicochemically and immunochemically characterized as free SP.

     

The half-lives of serum immunoglobulins in adult mice

We determined the half-lives of several sets of murine monoclonal antibodies spanning all immunoglobulin isotypes in the serum. The antibodies in each set possess the same V region. With this approach, the differences in half-life observed between the different isotypes are independent of the V region carried by the monoclonal antibodies and therefore must relate to each other in the same way as the half-lives of each class of serum immunoglobulins. The half-life of a monoclonal antibody of the gamma 2a isotype is identical to the average half-life of serum IgG2a as previously determined (6-8 days; P. Vieira and K. Rajewsky, Eur. J. Immunol. 1986. 16:871). Therefore, the half-lives determined with monoclonal antibodies possessing the same V region represent the half-life of the serum immunoglobulins. In this way we calculated the half-life of IgM as 2 days, IgG3 and IgG1 as 6-8 days, IgG2b has a half-life of 4-6 days. IgE has a half-life of 12 h. A polymeric form of IgA was found to be eliminated from the serum with a half-life of 17-22 h.     

Cross-reactivity of polyclonal serum antibodies generated against Cryptosporidium parvum oocysts.

Polyclonal antibodies raised against Cryptosporidium parvum oocysts were found to cross-react with Eimeria spp. oocyst antigens in an indirect immunofluorescence assay, and sera from Eimeria spp.-infected lambs reacted with some antigens from sonicated C. parvum oocysts (between 29 to 30 and 66 to 69 kDa) by Western blot (immunoblot). No cross-reaction was observed with cystozoites of Toxoplasma and Sarcocystis spp. These results show the existence of epitopes common to C. parvum and various Eimeria spp.     

Indirect immunofluorescence assay for the detection of hepatitis A virus-specific serum immunoglobulins.

Hepatitis A virus-specific BSC-1 cells were used for the detection of serum immunoglobulins to hepatitis A virus by indirect immunofluorescence. Of 150 serum samples tested, specific immunoglobulin M was detected only in patients with serologically confirmed acute hepatitis A, while specific immunoglobulin G was detected in patients with acute or past clinical hepatitis A as well as many patients with no known history of hepatitis.     

Immunochemical determination of gentamicin in serum. II. Preparation of polyclonal gentamicin antibodies]

The author describes the preparation of polyclonal rabbit antibodies against gentamicin. As immunogens gentamicin conjugates with bovine serum albumin were used, or else with thyroglobulin and haemocyanin. The immunization pattern involved combined administration of immunogens by the intravenous route, or by the intramuscular or intradermal route, using liposomes and complete Freund adjuvant. The highest antibody titres against gentamicin found by the ELISA method were obtained when procedures were used where the carrier was serum albumin and thyroglobulin and as adjuvant complete Freund adjuvant was used.     

In vivo effects of antiserum to IgD on surface immunoglobulins, serum immunoglobulins and lymphocyte blastogenesis in rhesus monkeys.

The effects of injecting monkeys with goat antiserum to IgD, the IgG fraction of that antiserum or normal goat serum (NGS) were compared. The subcutaneous injection of 4 ml/kg of the whole antiserum resulted in decreased percentages of lymphocytes with surface IgD or IgM lasting from day 1 through day 7 post-injection followed by substantial recovery on day 10. Lymphocytes from these animals were stimulated as indicated by the increased incorporation of 3H-TdR by cells placed in culture on days 7-21 post-injection. The increased blastogenesis occurred in rosette-depleted (B cell) populations and did not occur in rosette-enriched (T cell) preparations. Hypergammaglobulinaemia and increased concentration of serum IgG were first detected on day 10 postinjection, maximal on day 14 and were in decline by day 18. Injection of 4 ml/kg NGS did not alter the percentages of lymphocytes with surface immunoglobulins, result in hypergammaglobulinaemia, or stimulate the degree of blastogenesis observed after anti-IgD. Injection of the IgG fraction of the antiserum resulted in decreased lymphocytes with surface immunoglobulins but did not stimulate hypergammaglobulinaemia or increase blastogenesis. Injection of one monkey with the IgG fraction of anti-IgD combined with NGS resulted in increased serum IgG and increased blastogenesis. Both antibody to IgD and multiple antigenic challenge appear to be required for these responses.     

Determination of serum cholesterol concentration in the presence of ascorbate.

Pretreatment of a serum or plasma sample with ascorbate oxidase removed interfering ascorbate and allowed the determination of cholesterol to be carried out by a current enzymatic cholesterol method available in kit form. The Cobas-Fara was programmed to carry out pretreatment of the sample with ascorbate oxidase before addition of the cholesterol colour reagent.     

Dinitrophenyl-reactive immunoglobulins in the serum of normal bowfin, Amia calva

The sera from unimmunized bowfin agglutinate a large variety of red cells. Although they precipitate DNP-BSA they manifest only slight agglutinating capacity for DNP-coated cells. 15S immunoglobulin isolated by DEAE-cellulose chromatography followed by Sephadex G-200 gel filtration possessed a high level of broad reactivity towards unmodified DNP-coated cells, whereas the 7S immunoglobulin isolated by this procedure was inactive. However, following precipitation of whole serum with DNP-BSA both molecules could be recovered in a form which demonstrated specificity for DNP, in that both precipitated DNP-BSA and agglutinated DNP-coated cells but not unmodified cells. The mechanism of activation of the 7S molecule is not known but the data suggest that this immunoglobulin is divalent.     

Experimental bovine trypanosomiasis. Changes in the catabolism of serum immunoglobulins and complement components in infected cattle.

The turnover of serum proteins of calves experimentally infected with Trypanosoma congolense was compared to that of normal uninfected cattle. All proteins examined had much increased catabolic rates in infected animals. In normal animals the average half-lives in days for each protein were: IgG1 17.4, IgG2 22.4, IgM 4.8, IgA 3.4, IgE 1.9, C1 5.6 and C3 2.9. In trypanosome infected cattle the average half-lives were IgG1 1.9, IgG2 1.7, IgM 0.9, IgA 1.2, IgE 0.9, C1 1.2 and C3 1.1 days.