ABSTRACT. Polymorphonuclear leukocyte (PMN) func-GSD type IB is characterized by the clinical features of glucosetion was investigated in two patients with glycogen storage 6-phosphatase deficiency (GSD type IA), including hepatomegdisease type IB and neutropenia. Glycogen storage disease aly, growth retardation, fasting hypoglycemia, and lactic acidosis, type IB was documented by liver biopsy and a normal but normal latent enzymatic activity in the liver (l,2). Biochemamount of latent glucose-6-phosphatase activity. Patient A ical studies have demonstrated a defect in the microsomal transhad stomatitis, skin infections, and septicemia; patient B port system for G6P in GSD type IB (2, 3). Recurrent infection had respiratory infections, periodontitis, and oral candidi-occurs more prominently in type IB, and neutropenia is another asis. Absolute neutrophil counts ranged from 114 to 25801 distinguishing feature of this type (4, 5). Studies of PMN migramm3. Diminished and delayed migration of PMN into a tion in a patient with GSD type IB and neutropenia indicated skin "window" occurred in B. Random and directed PMN impaired PMN mobilization in vivo and impaired random and migration under agarose toward f-Met-Leu-Phe, pepstatin directed PMN migration in vivo (I). We have further investigated A, and zymosan-activated serum were severely diminished PMN function in two patients and demonstrated abnormalities in both patients. At lo-' M f-Met-Leu-Phe, mean random in metabolic and bactericidal functions as well as motility.and directed migration were 52 and 23% (A, n = 3) and 48 and 13% (B, n = 4) of controls. These results were inde-CASE REPORTS
An indirect double-antibody enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of human immunoglobulin E (IgE) and IgG to the cow's milk proteins (CMP) a-casein, a-lactalbumin, and P-lactoglobulin. Human serum albumin was used as the negative-antigen control. Rabbit anti-human IgE or IgG served as the primary antibody, and horseradish peroxidase-conjugated swine anti-rabbit immunoglobulin served as the secondary antibody. Positive control sera were obtained from patients with welldocumented histories of cow's milk allergy, while negative control sera were obtained from cord bloods of healthy full-term infants and from normal adult volunteers without known milk allergy. Test sera were obtained from 41 children (ages, 3 months to 13 years; average age, 2.6 years) with suspected cow's milk allergy and clinical manifestations that included wheezing, rhinitis, atopic dermatitis, urticaria, or gastrointestinal disturbances. The patients were simultaneously evaluated by prick skin testing with scratch test antigen to whole CMP. Although only 13 (32%) of the 41 patients were positive by the prick skin test, 25 (61%) were positive by the IgE ELISA. Of the 25 IgE ELISA-positive patients, 20 were also positive by the IgG ELISA. There was concordance of positive results between skin testing and the IgE ELISA in only 9 patients (22%), and there was concordance of negative results in 12 patients (29%). Discordant results were observed in 20 patients (49%). These results indicate that the ELISA is more sensitive than prick skin testing in the identification of individuals with elevated levels of IgE to CMP. Cow's milk supplementation in infant nutrition is now commonplace and is reflected in increased incidences of cow's milk protein (CMP)-induced adverse reactions which have diverse clinical manifestations (1, 13, 14). The lack of practical, efficient, and definitive diagnostic tests has restricted the identification of the allergy in these patients. Until recently, the techniques for the evaluation of suspected immediate-type hypersensitivity to CMP have been limited to skin testing, the radioallergosorbent test (RAST), and elimination-challenge tests, all of which are either impractical or lacking in both sensitivity and specificity. Enzyme-linked immunoadsorbent assays (ELISAs) are equal to RAST in sensitivity in the detection of allergen-specific immunoglobulin (IgE). ELISAs have several major advantages over RAST, including a long shelf life of reagents, nonradioisotopic probes, and inexpensive instrumentation requirements. These advantages make possible allergenspecific IgE testing in even the most modestly equipped laboratories. We have developed an indirect double-antibody ELISA for the detection of CMP-specific human IgE and IgG. All reagents used in the assay are commercially available, which simplifies their use in routine clinical laboratories. The procedure uses purified CMPs, i.e., ac-casein, oa-lactalbumin, and 3-lactoglobulin, which have been previously described
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