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Hyper immunoglobulin M immunodeficiency. (Dysgammaglobulinemia). Presence of immunoglobulin M-secreting plasmacytoid cells in peripheral blood and failure of immunoglobulin M-immunoglobulin G switch in B-cell differentiation.

The peripheral blood lymphocytes of nine patients with hyper immunoglobulin (Ig)M immunodeficiency were studied in an attempt to define the cellular basis of this disorder. B cells were normal in number but qualitatively abnormal in all patients. Approximately one-half of the B cell consisted of small lymphocytes (7-9 mum in diameter) bearing surface IgM and IgD, as well as C3 receptors. These cells were driven to secrete IgM but not IgG after in vitro stimulation by pokeweed mitogen. In the blood there were also large lymphocytes (10-14 mum in diameter) that possessed surface as well as intracytoplasmic IgM but lacked C3 receptors. These cells spontaneously secreted large amounts of IgM in vitro and on electron microscopy were found to be rich in rough endoplasmic reticulum. Such a subpopulation of lymphoid cells was not detected in normal peripheral blood and was unique for all patients with hyper IgM immunodeficiency studied.T cells from all patients were normal in number and in function both in vivo and in vitro and were able to generate adequate T-cell help to support IgG synthesis by normal B cells. No evidence was obtained for T cells capable of suppressing normal IgG synthesis in any of the patients after coculture with normal peripheral blood lymphocytes. The defect in hyper IgM immunodeficiency is intrinsic to B cells, which fail to switch from IgM to IgG synthesis.

Adult

Structural studies of the Xenopus 19S immunoglobulin and 7S immunoglobulin and two immunoglobulin-like proteins.

Xenopus laevis 19S and 7S immunoglobulins (Ig) were extensively reduced and alkylated, their H and L chains spearated and their molecular weights determined. Two kinds of L chains of molecular weight 25,000 and 27,000 were revealed by SDS-polyacrylamide gel electrophoresis. In addition two Ig-like proteins consisting of heavy chains only, of 19S H-type and with similar molecular weight, were detected in Xenopus serum ans isolated. These proteins share common antigenic determinants with Xenopus 19S Ig heavy chains and are devoid of light chain determinants.

Alkylation

Immunoglobulins and anti-Marek's disease virus antibody synthesis in chickens after passive immunization with immunoglobulin Y anti-Marek's disease virus antibody.

The effect of passive immunization with immunoglobulin Y (IgY) antibody against Marek's disease virus (MDV) was examined in MDV-susceptible chickens. The production of IgY, immunoglobulin M, and probably also immunoglobulin A was depressed in passively immunized chickens when compared with that in MDV-exposed chickens which had not been given IgY anti-MDV antibody. In passively immunized chickens, the synthesis of immunoglobulin M and IgY anti-MDV antibodies in response to MDV infection also was delayed as determined by agar gel precipitin and indirect fluorescence antibody tests.

Animals

[The diagnostic significance of immunoglobulin determination in chronic liver diseases. I. Differential diagnostic accuracy of immunoglobulin changes and enzyme activities (author's transl)].

715 patients with hepatobiliary diseases came for examination, who because of the morphological findings are classified in diagnostic groups. The immunoglobulin measurements of IGG, IGA, IGM were determined and assessed in combination with enzymatic investigations of GOT, GPT, AP and GGTP. Typical group-specific changes were only found in primary biliary liver diseases and toxic cirrhoses of the liver, all acute or chronic inflammatory liver diseases could not be separated and by inference from the final diagnosis showed great errors in classification. Data on the raised mean levels of immunoglobulins in the individual diagnostic groups were demonstrable, but the range limits were so widely scattered that their differential diagnostic valence is of no consequence. Immunoglobulins may appear to be of interest for the observation of the course of liver diseases, but they are unsuitable for diagnostic purposes.

Acute Disease

[Design of stable immunoglobulin erythrocyte diagnostica. 1. Erythocyte fixation and their sensitization by specific immunoglobulins].

The work presents the results of developing the method of fixation of erythrocyte constituting the cellular base of immunoglobulin erythrocytic diagnostic preparations and the sensitization of erythrocytes with immunoglobulin preparations of various specificity. Based on Ingraham's method, modified method of erythrocyte stabilization has been developed; it consists in the treatment of 50% cell suspension with 4% formaldehyde solution in the presence of 0.5% sucrose (erythrocyte suspension and formaldehyde solution being in the ratio 1 : 2.5). An economic and highly productive technique of sensitizing erythrocytes with immunoglobulin preparations has been developed. The essence of this technique lies in the interaction between 6% suspension of erythrocytes treated with formalin and tannin and the equal volume of sensitin taken in a working dose. The work also presents the method of synthesizing the bifunctional compound fluoro-borate bis-daizonium complex (obtained from benzidine) and discusses the comparative possibilities of the methods of developing immunoglobulin erythrocytic diagnostic preparations by sensitization of tannin-treated erythrocytes and by chemical conjugation.

Allergens

Heterogeneity of nonimmune immunoglobulin Fc reactivity among gram-positive cocci: description of three major types of receptors for human immunoglobulin G.

Two hundred and thirty strains of various gram-positive cocci were tested for quantitative, nonimmune binding of radiolabeled human polyclonal immunoglobulin G (IgG). The majority of coagulase-positive staphylococci and streptococci belonging to serogroups C and G showed a high uptake of IgG. The binding of immunoglobulin to group A streptococci was considerably less, with a number of strains completely negative. None of the pneumococcal or the group B or D streptococcal strains displayed any binding capacity. Heterogeneity of the IgG reactivity of various reactive strains was studied in an inhibition assay using 10 different animal serum pools. Three different inhibition patterns were seen, each of them revealing a striking degree of homogeneity within single bacterial species. Staphylococcus aureus and group A streptococci, respectively, constituted two homogeneous groups which differed markedly from each other and from C and G streptococci. No differences were observed between group C and G streptococci. Based on the profound differences between these homogeneous groups, three major types of Fc receptors could be defined. Type I and II Fc receptors were found on S. aureus and on group A streptococci, respectively. Fc receptor type III represented the immunoglobulin-binding structure of both group C and G streptococci.

Animals

[Immunoglobulins in swine. (B) Immunohistological studies on the localization of cells containing immunoglobulin in the tissues of the small intestine and spleen (author's transl)].

Antisera to porcine immunoglobulins, conjugated with flurescein isothiocyanate, were used in studies on the localization of cells containing immunoglobulin in the tissues of the intestine and spleen in pigs. Cells containing IgM and IgA were found to be present in the lamina propria of the intestinal mucosa, the villous membrane showing fluorescence due to liberated immunoglobulins. Cells containing IgG and IgM were detected in the marginal areas of the splenic lymph nodes but could not be definitely identified as mature plasma cells.

Animals

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.

Animals

Loss of antibody activity in human immunoglobulin A exposed extracellular immunoglobulin A proteases of Neisseria gonorrhoeae and Streptococcus sanguis.

Immunoglobulin A (IgA) proteases are extracellular enzymes elaborated by Neisseria gonorrhoeae, N. meningitidis, and Streptococcus sanguis. These enzymes each cleave human IgA1 at a critically situated prolyl-threonyl peptide bond to yield Fab alpha and Fc alpha fragments. To study their effect on the antibody activity of human IgA, we enzymatically digested a group of five human IgA monoclonal immunoglobulins with high-titer rheumatoid factor or cold agglutinin activity and human serum macroamylase, an amylase-IgA complex. In contrast to four control IgM rheumatoid factor monoclonal proteins, whose activity was unaffected by enzyme, gonococcal and streptococcal IgA proteases caused prompt, major reductions of IgA antibody activity to negligible levels and converted macroamylase activity to amylase of normal size, as determined by molecular sieve chromatography. In addition, both enzymes promptly deagglutinated sensitized cells that had been aggregated by IgA rheumatoid factors, indicating that IgA bound to antigen is also susceptible to enzyme cleavage. Fab fragments of Iga protein Chr, a rheumatoid factor, showed essentially no antigen-binding activity despite the high titers observed with the parent protein. These studies emphasize the high degree of specificity of the microbial proteases for IgA and their potential for interfering with antibody activity in the IgA1 subclass.

Agglutination

The effects of anti-IgD on serum immunoglobulins, antibody production, and immunoglobulin-bearing cells in adult rats.

Rabbit anti-rat IgC (delta-specific) injected subcutaneously into adult rats resulted in hypergammaglobulinemia, increased serum immunoglobulins, increased antibody production, and alterations in the percentages of PBL expressing IgD and IgM. Serum IgM, IgG2a and IgG2c concentrations were increased 100%, 125%, and 200%, respectively, over controls, whereas IgA levels were not altered. Increases in the serum immunoglobulins were predominately due to an increase in antibodies to the rabbit serum components. A depletion of IgD-IgM bearing lymphocytes in the peripheral blood was observed after 24 hr of injection. Percentages of IgC-IgM bearing lymphocytes returned to normal by day 4 and were significantly increased (approximately 2-fold) by day 8. The in vivo effects of anti-delta in rats were similar to those reported in the monkey. Possible modes of action of the anti-delta and the significance of these results in relation to the biologic function of membrane IgD are discussed.

Animals

Sequential expression of immunoglobulin on developing mouse B lymphocytes: a systematic survey that suggests a model for the generation of immunoglobulin isotype diversity.

Paired immunofluorescent staining with antibodies specific for the major isotypes of mouse immunoglobulin was used to study the ontogenetic expression of diversity of cell surface immunoglobulin. The first B lymphocytes to emerge, derived from cytoplasmic IgM+ precursors, express sIgM exclusively. Between birth and 3 days of age separate populations of sIgM+ B lymphocyte acquire a second isotype: sIgD, one of the subclasses of sIgG, or sIgA. At 3 days, all splenic B lymphocytes that bear sIg or sIgA also express sIgM, but virtually none stain for sIgD. By 7 days, a substantial porportion of sIgG+ or sIgA+ lymphocytes in spleen and most of those in lymph node express both sIgM ans sIgD. Anti-mu antibody treatment from birth prevented development of B lymphocytes expressing any isotype. These observations suggest that the immature sIgM+ B lymphocyte is the pivotal cell in the generation of the different sublines of B cells and that sIgD ig or IgA. The frequency of lymphocytes bearing only sIgG or sIgA is higher in old than in young mice, suggesting that sIgD and sIgM may be lost after stimulation by antigens. The occurrence of a nearly identical distribution of sIg isotypes on B lymphocytes from athymic, pathogen-free mice suggests that primary expression of isotype diversity does not require T cells.

Aging

Murine plasma cells secreting more than one class of immunoglobulin heavy chain. III. Immunoglobulin production by established cultures and cloned lines of SAMM 368.

SAMM 368, a plasmacytoma which produces IgA-kappa and IgG2b-kappa was established in vitro from primary explants or after animal passage. The 9 lines that were established produced both paraproteins. The production of both immunoglobulins by single cells was demonstrated by immunofluorescent staining and cloning. Continuous culture of 3 parent lines for 18 months and 11 cloned lines for periods from 5 to 7 months demonstrated that double production is a stable characteristic of this plasmacytoma. Two single paraprotein-producing varients (IgG2b or IgA) were derived when cells were cultured in the presence of Fungizone. Chromosomal analysis of SAMM 368 indicates that this double producing tumor has a modal number similar to those observed in myelomas producing a single immunoglobulin class.

Cell Line

Immunologic aspects of lepromatous leprosy as related to the immunoglobulins of external secretions: salivary immunoglobulins.

A significant reduction in salivary immunoglobulins in lepromatous leprosy is recorded as compared to normal subjects. Saliva of undernourished subjects and patients with oral malignant tumors, which were studied as controls, showed an appreciable rise in IgA levels in both. It is suggested that impairment of T cell function, which is associated with lepromatous leprosy, is not responsible for the observed low level of salivary immunoglobulins in lepromatous leprosy.

Adult

Mouse lymphocytes with and without surface immunoglobulin: preparative scale separation in polystyrene tissue culture dishes coated with specifically purified anti-immunoglobulin.

Mouse spleen cells could be preparatively separated into immunoglobulin positive (Ig+) and immunoglobulin-netative (Ig-)populations by incubating as many as 2 X 10(8) cells per 100 mm diameter petri plate coated with specifically purified goat anti-mouse immunoglobulin. The non-adherent population was 95% or more Ig-, and possessed graft versus host and cytotoxic effector activities, as would be expected for T cells. They could also give a mixed lymphocyte reaction and generate cytotoxic effector activity on culture in vitro. The adherent cells could not be released undamaged from plates coated with undiluted anti-Ig, but they could be released from plates coated with a 1/4 or 1/10 dilution of anti-Ig in an irrelevant antibody. The released cells were over 90% viable by trypan-blue staining, and 94% or more of the viable cells were Ig+.

Animals

Induced synthesis of immunoglobulin messenger RNA accompanies induction of immunoglobulin production in cultured mouse spleen cells.

Immunoglobulin kappa type light chain mRNA (Lkappa mRNA) accumulated in parallel with secretion of immunoglobulin M in cultured mouse spleen cells activated by lipopolysaccharide. Actinomycin D suppressed the accumulation of kappa chain mRNA completely without affecting the degradation rate of kappa chain mRNA. The half life of kappa chain mRNA was about 9 h. Available evidence indicates that lipopolysaccharide stimulates de novo synthesis of kappa chain mRNA. The accumulation of kappa chain mRNA was markedly suppressed by inhibitors of DNA or protein synthesis such as hydroxyurea, cytosine arabinoside and cycloheximide.

Animals

Practical methods of determining serum immunoglobulin M and immunoglobulin G concentrations in foals.

Serum concentrations of immunoglobulin M (IgM) and immunoglobulin G (IgG) can be determined in the horse with a satisfactory degree of accuracy, using commercially available reagents. Selected lots of anti-human IgM can be used in precipitation tests to detect and quantitate equine IgM. Commercially available anti-equine IgG tended to overestimate the amount of IgG in single radial immunodiffusion tests. Even with these limitations, commercial reagents can be used to differentiate immunodeficiency disorders of foals, including combined immunodeficiency and failure of passive transfer of colostral antibody from mare to foal.

Animals