Binding between components of the tubercle bacillus and humoral antibodies.

Studies were undertaken to find a substance or substances for use in primary binding types of tests to detect humoral antibodies in rabbits and monkeys exposed to the tubercle bacillus that would distinguish between strains of mycobacteria. The antigen employed was a component of the 5159 strain of Mycobacterium tuberculosis that was obtained following sonification, ultracentrifugation, electrophoresis, and elution from a preparative polyacrylamide column. When the antigen was labeled with (131)I, specific binding was observed in sera from immunized rabbits and BCG-protected rhesus monkeys by the radio-gel electrophoresis and ammonium sulfate tests. This component was partially characterized, and its major antigenic determinants were associated with anionic mucopolysaccharides. Electrophoretically at pH 8.3 it migrated anodally to albumin, its molecular weight was between 9,000 and 12,000, and it was soluble in 50% saturated ammonium sulfate. Binding to antibody was destroyed after treatment with Pronase, but not after DNase or RNase. Inhibition of the reaction, as measured by the ammonium sulfate test, between the (131)I-labeled component and antisera from rabbits that had been immunized with sonicated 5159 organisms, was studied. These experiments demonstrated a capacity to define subtle similarities and differences among different mycobacteria and mycobacterial components. Some microorganisms not clearly related to mycobacteria also partially inhibited this reaction, suggesting that they shared antigenic groups with the component derived from 5159 organisms. The studies described suggested the advisability of using direct primary tests and purified components of mycobacteria to differentiate further the antigenic groups between individual pathogenic mycobacteria and between pathogenic and nonpathogenic organisms.

Reaginic activity associated with IgG immunoglobulin.

Five human sera with reaginic activity to a number of allergens were fractionated using anion exchange chromatography. In each serum, fractions which contained detectable IgG and no detectable IgA had capacity to fix to skin and subsequently elicit a P-K reaction. Four of these sera had reaginic activity about equally distributed between fractions containing only IgG and fractions containing mixtures of IgG and IgA. A fifth serum contained reaginic activity to crystalline bovine serum albumin (BSA) and most of the activity was associated with the fraction which contained only IgG. This serum was extensively studied using a variety of techniques and it was confirmed that most of the reagin to BSA in this serum was in those fractions containing only IgG. Since reaginic activity can no longer be considered a unique property of IgA the implications of finding antibody with reaginic qualities in immunoglobulins other than IgA are discussed.

Shared antigens between Mycobacterium bovis (BCG) and other bacterial species.

Antigens from Mycobacterium bovis and Listeria monocytogenes bind to serums from normal rabbits as well as from those immunized with unrelated bacteria, especially intracellular parasites. Binding is to the Fab portion of immunoglobulin G and is immunologically specific.

Suppression and immunotherapy of the guinea pig line-10 hepatocarcinoma mediated by nonviable Staphylococcus aureus.

Suppression of growth of the line-10 guinea pig hepatocarcinoma was achieved after the simultaneous injection of line-10 cells and heat-killed Staphylococcus aureus. Growth of tumor was also suppressed when line-10 cells were injected alone, contralaterally, at the same time as the vaccine mixture. Immunity developed to subsequent line-10 cell challenges but not to other syngeneic tumors. Similar results were obtained with the use of protein A-rich or -deficient strains of S. aureus. Multiple intratumor injections of heat-killed S. aureus were therapeutically effective against 6-day tumors. The antitumor effects of nonviable S. aureus were similar in many ways to those of living BCG or bacterial products suspended in oil droplets.

Prevention of tumor growth after intradermal injection of BCG extracts: a comparison of results in strain-2 guinea pigs from the National Institutes of Health and from the National Jewish Hospital and Research Center.

Line 10, a transplantable hepatocellular carcinoma, was obtained originally from an NIH strain-2 male guinea pig fed diethylnitrosamine. The antitumor activity of BCG and BCG extracts was evaluated in strain-2 guinea pigs obtained both from NIH and the National Jewish Hospital and Research Center (NJH). Animals were immunized with these materials and then tested for their capacity to resist the growth of intradermally injected line-10 tumor cells. Tumor growth was not prevented in 18 NIH animals immunized with living BCG. No tumor growth occurred in 1 of 22 NIH animals immunized with a residue that remained after exhaustive methanol extraction of BCG and in 1 of 44 NIH guinea pigs immunized with BCG extracts. In contrast, tumor growth was prevented in 13 of 22 similarly immunized NJH guinea pigs.

Transmission of BCG-associated tumor resistance from maternal to newborn guinea pigs.

Resistance to line-10 hepatocarcinoma was studied in the newborn offspring of inbred strain-2 Sewall Wright guinea pigs that had recovered from this tumor after prior administration of subcellular components of Mycobacterium bovis strain BCG. These mothers gave birth to 15 offspring, 53% of which resisted repeated challenges with this tumor. Offspring of mothers who had been given only BCG components were similarly tested. Complete protection was observed in 23.5% of 17 babies and attenuation of disease in many others. All control offspring from normal, untreated mothers succumbed to this tumor. Resistance was associated with delayed cutaneous hypersensitivity reactions to tumor and mycobacterial antigens.

Enhanced immunogenicity of line 10 guinea pig hepatocarcinoma cells after culture.

Line 10 hepatocarcinoma cells derived from ascites in a strain 2 guinea pig were tumorigenic when transferred intradermally. After they had been cultured in vitro for 20 days or more in medium enriched with 10% fetal bovine serum (FBS), they became immunogenic. Injections of immunogenic cells did not cause lethal tumors, and recipients were resistant to subsequent challenges with tumorigenic line 10 cells. Resistance was specific since growth of line 1 cells, a syngeneic but antigenically distinct tumor, was not affected. Cells cultured in medium enriched with 10% calf bovine serum or 10% normal guinea pig serum or in reduced concentrations of FBS were less effective in inducing resistance. When cultured line 10 cells were injected ip into normal guinea pigs, ascites tumors developed that were tumorigenic. The growth rate of line 10 cells in culture was considerably decreased as determined by reduced [3H]thymidine incorporation and mitotic indices. The mechanism(s) responsible for enhancement of immunogenicity in cultured line 10 cells is discussed but was not determined.

Comparison between murine natural antibodies and natural killer cells: recognition of separate target structures as revealed by differential in vitro expression and dependence on glycosylation.

The specificity of complement-fixing, cytotoxic antibodies against the YAC lymphoma in sera of normal young adult (A X C57BL)F1 mice was studied. In vivo-maintained, immunoselected sublines of the YAC lymphoma expressed low amounts of the natural antibody (NAb) target structure. These cell lines were also resistant to natural killer (NK) cell-mediated lysis. After 2-3 weeks of in vitro culture the immunoselected cell lines became NK sensitive, but they remained resistant to NAb. When several independently derived variants selected for low NK sensitivity were tested for their ability to absorb NAb, the degree of absorption varied considerably among the variants. NAb could be inhibited by purified C-type virus particles and also by bacterial sonicates and various glycoprotein preparations. Treatment of target cells with tunicamycin, an inhibitor of asparagine-linked glycosylation, decreased the sensitivity to NAb lysis but had no impact on NK sensitivity. Thus the results indicated that a) NAb and NK cells recognized separate target structures and b) the target structure(s) for NAb but not for NK cells were saccharides of the N-glycosidic type.

Suppression and immunotherapy of the guinea pig line 10 hepatocarcinoma mediated by heat-killed disrupted Mycobacterium bovis strain Bacillus Calmette-Guérin.

Heat-killed Mycobacterium bovis strain Bacillus Calmette-Guérin cells were sonically disrupted, and their antitumor effects against the line 10 hepatocarcinoma in strain 2 guinea pigs were evaluated. When injected together with viable line 10 cells, there was complete suppression of tumor growth. Growth of tumor was also suppressed when line 10 cells were injected contralaterally at the same time as the vaccine mixture. Multiple intratumor injections of sufficient disrupted M. bovis strain B. Calmette-Guérin were therapeutically effective against 74% of 7-day-old tumors and against 40% of 14-day-old tumors. Surviving animals were usually resistant to subsequent rechallenges with line 10 cells but not to syngeneic L2C leukemia cells. By means of a competitive radioimmunoassay, antigenic determinants were detected that were expressed by disrupted but not by intact bacteria.

Shared antigens between bacteria and guinea pig line 10 hepatocarcinoma cells.

This study was undertaken to investigate the possibility that Listeria monocytogenes, Brucella abortus, and Salmonella typhimurium share antigenic components with guinea pig line 10 hepatocarcinoma cells. Rabbits were immunized with sonicates of these bacteria or line 10 tumor cells. Other rabbits were immunized with line 1 cells, a tumor with antigenic characteristics different from those of line 10. The binding of antibodies to radiolabeled antigens prepared from extracts of bacteria and line 10 cells was studied by precipitation of radiolabeled antigen-antibody complexes with anti-rabbit immunoglobulin. Antibodies in sera from rabbits immunized with these bacteria and line 10 cells bound both the labeled bacteria and line 10 antigens. Antibodies in sera from rabbits immunized with line 1 cells did not bind the bacterial antigens. Inhibition studies involving reactions between radiolabeled Listeria and line 10 antigens and antibodies to Listeria and line 10 cells confirmed that the binding reactions were specific and that line 10 cells shared antigens with Listeria cells. The possibility that B. abortus and S. typhimurium also shared antigens with line 10 cells was suggested. Whether antigens shared by these bacteria and line 10 cells are identical with tumor-specific antigens was not determined.

Immunotherapy of the guinea pig line 10 hepatocarcinoma with a variety of nonviable bacteria.

A variety of heat-killed bacteria were tested for their capacity to induce regressions of established line 10 hepatocarcinomas in syngeneic guinea pigs. Multiple intralesional injections of heat-killed Escherichia coli, Streptococcus mutans, Listeria monocytogenes, and Propionibacterium acnes resulted in complete regression of the tumor in a majority of guinea pigs. Repeated injections of heat-killed Mycobacterium bovis strain Bacillus Calmette-Guérin caused no regressions. Surviving animals were immune to subsequent challenge with line 10 cells but not L2C cells, another syngeneic tumor.

Purification of antibodies to bacterial antigens by an immunoadsorbent and a method to quantify their reaction with insoluble bacterial targets.

A combination of procedures was employed to develop a radioimmunoassay which quantified the binding of antibodies to antigens of either intact Propionibacterium acnes or to antigens of insoluble extracts derived from the bacteria. Reactive antibody populations were purified by use of bacterial immunoadsorbents which were prepared by coupling P. acnes to diethylaminoethyl cellulose. Binding of antibodies was detected with [125I]staphylococcal protein A ([125I]SpA) and optimal conditions for the assay defined by varying the amounts of antibodies, bacterial antigenic targets and [125I]SpA. In antibody excess, 100% of available [125I]SpA was bound by the target-antibody complexes. However, when antibody concentration was limiting, a linear relationship was demonstrated between per cent specific binding of [125I]SpA and antibodies bound to bacterial targets. These results were achieved only with immunoadsorbent-purified antibody populations and not with hyperimmune sera or IgG. The radioimmunoassay detected subtle antigenic differences and similarities between P. acnes, P. acnes extracts and a variety of unrelated microorganisms.

Humor as the focal point of treatment for forensic psychiatric patients.

Framed as a qualitative case study based on constructivist notions, the research being reported evaluates the Humor Group, a unique treatment modality evoking therapeutic change by engaging male forensic psychiatric patients and nursing students in humorous activities. The group's meaning for patient health was investigated by cross-analyzing data collected from patient and student questionnaires, in-depth patient interviews, and naturalistic observations made by the researcher during the group's 4-year tenure. The Humor Group structure and format are offered as guides for undertaking similar attempts to rein in humor's healing potential in other settings and with other populations.

The Victim Care Service: a program for victims of sexual assault.

Sexual victimization is a singularly traumatic event. At times, however, the implicit danger of the trauma experience itself gets exaggerated. For the victim the more lasting damage frequently stems from how he or she was treated by friends, family, health care providers, and public authorities following the incident. Treating the victimized patient as quickly and sensitively as possible is essential to minimizing the potential for further victimization. The Emergency Department at Boston City Hospital is frequently the site for the acute management of sexual assault victims. The Victim Care Service is designed to prevent secondary traumatization of such victims. The five-phase program and the fundamental assumptions upon which the phases are based are presented.