Diagnostic methods to demonstrate igE antibodies: skin testing techniques.
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Biomedical subjects
Publications and source records attributed to W T Kniker.
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Five patients received ketoconazole treatment of chronic mucocutaneous candidiasis. One also had disseminated histoplasmosis and cryptococcosis. Ketoconazole was well absorbed after an oral dose of 200 mg and produced detectable antifungal blood levels for more than eight hours after each dose. Thrush cleared in less than two weeks in all patients. Cutaneous lesions cleared slowly during several months of treatment. A patient with polymycotic infection had a rapid clearing of candidiasis, but her histoplasmosis did not respond to three weeks of treatment. Adverse drug effects were minimal. Ketoconazole is a promising new agent for treatment of chronic mucocutaneous candidiasis.
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Functions of polymorphonuclear leukocytes including chemotaxis, phagocytosis, and microbial killing were investigated in baboons (Papio cynocephalus), cebus monkeys (Cebus apella), and marmosets (Saguinus oedipus). Cells from a group of normal adult human volunteers were also studied for comparison. Polymorphonuclear leukocytes from the three nonhuman primate species were comparable to each other and to humans in chemotactic activity using endotoxin activated serum. In addition, the ability of blood leukocytes to ingest and kill microorganisms was investigated using a new fluorochrome microassay. Leukocytes of all non-human primates were competent in phagocytosing bacteria, but leukocytes from cebus monkeys and marmosets ingested candida organisms poorly. Compared to humans, all nonhuman primates exhibited poor killing of the microbes.
While delayed cutaneous hypersensitivity (DCH) testing with ubiquitous antigens is acknowledged to be useful in assessment of cell mediated immunity, existing methods suffer from lack of standardized antigens and optimal doses. The authors have evaluated a new plastic, disposable device (Multi-Test, Lincoln Laboratories) for simultaneous administration of up to eight test materials by multiple puncture and seven ubiquitous antigens in 70% glycerol prepared by Institut Merieux. Each antigen was standardized chemically and biologically for DCH activity. The Multi-Test system proved to be safe, rapid, painless, and reproducible means of evaluating DCH to multiple standardized antigens. This procedure offers the potential for detecting changes in cell-mediated immunity in diseases individuals who are skin tested periodically.
The Multi-Test applicator is a disposable plastic device for the simultaneous prick-puncture administration of different antigens at eight sites, in a rapid, relatively painless and reproducible manner. To determine its utility in immediate or reaginic skin testing, it was compared with the intradermal technique in children and adults. Skin test responses to glycerinated inhalant antigens, 1:20 or 1:10 W/V, administered by Multi-Test were comparable to those obtained from the same antigens applied intradermally in aqueous dilutions of 1:1,000 or 1:1,500 W/V.
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Using skin test response and lymphocyte blastogenesis as indicators of cell-mediated immunity, we have been able to demonstrate in vivo transfer of cell-mediated immunity to marmosets both with dialyzable transfer factor (TFd) prepared from a human donor and transfer factor (TF) from baboon whole cell lysates. We were able to protect marmosets with TFd from fatal Herpesvirus hominis type 1 (HVH-1) disease. When TFd was administered prior to challenge with HVH-1, 50% of the marmosets survived. Of the untreated control animals and those first treated on day 0 or postinfection, 100% succumbed to disseminated HVH-1 disease. In addition, when TF prepared from TFd-treated marmosets which had survived HVH-1 disease, was given to other marmosets, it conferred protection from subsequent HVH-1 challenge.
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One-way mixed lymphocyte culture (MLC) reactivity and skin graft rejection were investigated to assess immune competence and histocompatibility in the baboon, the cebus monkey, and the cotton-topped marmoset. Adequate and comparable cell-mediated immunity was demonstrated. MLN reactions revealed strong histocompatibility differences among the three primate species and stimulating alloantigens for unrelated individuals of each species. Xenogeneic skin grafts survived nine days and allogeneic grafts, an average of 11 days. These systems appear readily applicable for assessing histocompatibility in nonhuman primates.
E (erythrocyte) and EAC (erythrocytes coated with antibody and complement) rosettes were quantitated in baboons, cebus monkeys and cotton-topped marmosets. There was poor correlation between E rosette levels and other parameter of T-lymphocyte function. In nonhuman primates, E rosettes were increased in the presence of thymosin fraction V, while human E rosettes were not affected.
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The present report describes an infant with severe combined immunodeficiency and cartilage-hair hypoplasia whose lymphocytes responded to thymosin in vitro. Immunologic evaluation was undertaken at 4 1/2 months of age following a history of recurrent severe infection. Family history included three cousins who died in early infancy, one from streptococcal meningitis and pneumonia, one from generalized varicella, and another from reticuloendotheliosis. Quantitative immunoglobulins were markedly depressed: IgG 141, IgA 0, and IgM 24 mg/100 ml. There was an absolute lymphopenia, multiple skin tests were negative, and in vitro lymphocyte responses to mitogens and antigens were depressed. Spontaneous E rosette determinations were 21% compared with control values of 65.7%. Erythrocyte adenosine deaminase (ADA) activity was normal. The patient's E rosette formation increased in the presence of thymosin, fraction 5, reaching a maximum of 56% with a concentration of 500 mug thymosin. Blastogenic responses to phytohemagglutinin also increased in the presence of thymosin. Transplantation of 24-week fetal thymus in Millipore diffusion chambers and subsequently transplantation of 18-week fetal thymus by intraperitoneal injection was accomplished. E rosettes increased to 35-40% and blastogenic responses to mitogens increased. Eight days after the second transplant the patient underwent a mild graft vs. host reaction which subsided after 1 week and mitogen blastogenic responses again increased to 5-8 times previous values, but still well below control ranges. Repeated episodes of pulmonary infection ensued, cor pulmonale resulted, and the clinical course was relentlessly downhill with the patient expiring from respiratory failure 5 months after transplantation.
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