Halina Borel

Selective Gamma-G Globulin Deficiencies in Patients with Recurrent Pyogenic Infections

Abstract Three patients with lifelong susceptibility to pyogenic infections and progressive pulmonary disease were found to have selective deficiencies in their γG globulins. The serum of the first patient contained abnormally low concentrations of γG1, γG2 and γG4; the serum of the second patient was deficient in γG1, γG2 and γG3, and the third patient lacked γG1 and γG2. The serum concentrations of the other immunoglobulins were either normal or elevated. None of these abnormalities could be shown to have a genetic basis. Gamma-globulin replacement therapy prevented recurrence of infection in these patients.

A novel technique to link either proteins or peptides to gammaglobulin to construct tolerogens.

In order to extend the concept of constructing tolerogens (i.e., compounds which induce immunologic tolerance), we developed a novel method to covalently link either protein or peptide to isologous gammaglobulin. We used disuccinimidyl suberate (DSS) for preparing protein conjugates in solution in a novel use of this reagent. We tested the efficacy of this method in two different experimental models: in the first, we found that administration of pigeon cytochrome C conjugated to mouse IgG in vivo induces T cell unresponsiveness in vitro. In the second, we induced unresponsiveness to factor VIII light chain both in newborn and, more importantly, in adult mice already immune to factor VIII. We hope that this simple method will provide a powerful tool to construct tolerogens useful in the specific treatment of either allergic or autoimmune diseases.

Conjugation of DNA fragments to protein carriers by glutaraldehyde: Immunogenicity of oligonucleotide-hemocyanin conjugates☆

The practical realization of the concept of specific immunotherapy for systemic lupus erythematosus (SLE) has been hampered, thus far, by an inability to link DNA fragments to carrier protein. In this paper, a novel technique is described, in which glutaraldehyde is the linking agent. A 2-stage method was used to link oligonucleotides to a soluble protein carrier, such as keyhole limpet hemocyanin (KLH) or human gamma globulin (HGG), whereas a 1-stage technique was sufficient to link oligonucleotides to sheep red cells. Both the ultraviolet absorbance spectrum and diphenylamine assay demonstrated that oligonucleotides were coupled to soluble protein. The conjugate of oligonucleotide to protein carrier appears to be recognized by anti-DNA antibody since oligonucleotide linked to either KLH or HGG inhibited the binding of anti-DNA antibody in vitro, and oligonucleotide-coupled sheep cells are agglutinating by seropositve sera from lupus patients. In addition, oligonucleotide-KLH raised hemagglutinating antibody to denatured DNA in C57BL/6, DBA/2 or NZB mice, as well as IgG antibody as detected by SPRIA in C57BL/6 and DBA/2 mice. The significance of this new method for the development of an antigen specific therapy of SLE is discussed.

Auto-immunosuppression: Recurrent infections associated with immunologic unresponsiveness in the presence of an auto-antibody to IgG☆

Abstract A 14-year-old French Canadian male with chronic progressive bronchiectasis and undue susceptibility to infection over a duration of 4 years was found to be hypogammaglobulinemic. His serum contained a high titer of IgM antibody to Fc fragment but it had no Gm specificity. This antibody was also lymphocytotoxic and its complement-dependent lympocytotoxicity was not HL-A associated and was blocked by a gamma G globulin or Fc fragment of gamma globulin. No B or T cell function could be measured in the patient.

Influence of intravenous administration of nucleoside-coupled spleen cells on murine lupus nephritis in female (NZB × NZW)F1 mice

In this study, we examine the influence of the intravenous administration of nucleoside-coupled spleen cells on the spontaneous development of murine lupus nephritis in young and adult BWF1 mice. In both age groups, this treatment failed to affect the autoimmune disease. Rather, in young mice administration of nucleoside-coupled spleen cells accelerates the appearance of anti-DNA antibody suggesting that BWF1 have a specific defect in the immunoregulation of anti-DNA antibody production. The significance of this finding for the pathogenesis of the disease in BWF1 mice is discussed.

Immunologic tolerance to DNA in B cell lines from both normal and autoimmune mice

SummaryWe examine whether B cell lines enriched for DNA specificity from either autoimmune (BWF1) or normal mice (Balb/c) can be rendered unresponsive to autoantigen in terms of the specific suppression of direct antibody-forming cells to DNA. These B cell lines were both Lyt-1 positive and negative. Preincubation with oligonucleotide, covalently linked to mouse γ-globulin, specifically suppressed the antigendriven response elicited by DNA horse red blood cells in B cell lines from both strains of mice. There is a 5-fold difference in susceptibility to DNA-specific tolerance induction between B cell lines of BWF1 and Balb/c mice. Thus, B cells from autoimmune mice do not appear to have an inherent absolute defect in being rendered tolerant to autoantigen, but are relatively less susceptible to DNA-specific tolerance than nonautoimmune cell lines.

A New Genetic Determinant of γG3

Abstract. A new antigenic determinant Ry confined to the γG3 subgroup is described. It was revealed by an antibody detected in the serum of a healthy Caucasian female. Ry is present in almost all normal human sera of all Gm phenotypes. However, its absence is associated with the Gmazg and Gmazb0b3st haplotypes and the absence of the Gm(f) determinant on the γG1 subgroup.

Detection of antibody-forming cells and humoral antibody to horseradish peroxidase

Two new simple methods for detecting antibody-forming cells by hemolytic plaque assay and hemagglutinating antibody to horseradish peroxidase have been developed in mice. Both techniques utilize as target, sheep erythrocytes coupled directly with horseradish peroxidase. These assays are sensitive, antigen-specific and are useful to quantitate both direct and indirect antibody-forming cells and humoral antibodies.