Daniel J. Marmer

Immunologic aspects of human endometriosis.

ABSTRACT: General and specific immune function were examined in women with endometriosis. Nonspecific parameters included total leukocyte and differential counts, quantitative immunoglobulin (IgG, IgA, and IgM) determinations, total hemolytic complement, C3, C4, mitogen‐induced lymphocyte stimulation, and human leukocyte antigen (HLA) typing; no differences were observed when data were compared to age‐matched controls without endometriosis. In contrast, the specific immune response T‐lymphocyte‐mediated cytotoxicity to autologous endometrial cells was significantly reduced (P <0.01) in women with endometriosis. When results from patients were analyzed according to clinical severity of endometriosis, even more striking immunologic alterations were delineated. In addition, lymphocyte stimulation responses to autologous endometrial antigen were lower in patients with severe or moderate disease, approaching statistical significance (P = 0.18 and 0.12, respectively). These studies suggest an immunologic basis for development of endometriosis.

Immune deficiency in fetal alcohol syndrome.

Summary: In a review of 13 documented cases of fetal alcohol syndrome (FAS), an increased incidence of life-threatening bacterial infections as well as a propensity to minor infections was observed. Five of 13 patients had had at least one episode of pneumonia, two had meningitis, and one had sepsis. A comprehensive immunologic evaluation of FAS was completed, and results were compared to an age-matched control group of children with intrauterine growth retardation without FAS.Children with FAS were shown to have decreased E rosette-forming lymphocytes (35 ± 5% versus 55 ± 5% or 1328 ± 274 versus 2333 ± 112 per mm3), low EAC rosette-forming lymphocytes (15 ± 2% versus 18 ± 1% or 524 ± 109 versus 740 ± 75 per mm3), and diminished mitogen-induced stimulation responses to mitogens: 31616 ± 5337 versus 58076 ± 4455 cpm for phytohemagglutinin, 17582 ± 5436 versus 35018 ± 5346 for pokeweed mitogen, and 32460 ± 7044 versus 54996 ± 5531 for concanavalin A, P < 0.05. Nine patients had dysgammaglobulinemia. FAS subjects also had a marked eosinophilia (624 ± 154 versus 72 ± 27 mm3). Other parameters of immune function including absolute lymphocyte and neutrophil counts, total hemolytic complement, delayed cutaneous hypersensitivity, and nitroblue tetrazolium dye reduction assays, were not different from control children. Impairment of immunity may explain an increased susceptibility to infection in FAS.Speculation: Defects in host defense and propensity to infection are attributable to intrauterine exposure to high levels of alcohol. Such abnormalities are not related to the degree of postnatal growth retardation nor to degree of malnutrition and do not correct with increasing age. Subsequent disease processes associated with defects in immunity such as malignancy or autoimmunity could result from this exposure to alcohol during fetal development.

Surgically induced immunologic alterations in the child

Surgery is generally believed to be an immunodepressant. This assumption is based, in part, upon studies of compromised patients undergoing major operation. Similar studies in normal adults following elective procedures are contradictory and little information is available regarding the pediatric surgical patient. This paper presents a study of immune function in children undergoing elective operation. Fifty healthy preoperative children (mean age: 20 months) were randomly selected. Ninety-five percent underwent inguinal herniorrhaphy. Operative time averaged 45 minutes (range: 30 to 90 minutes). Anesthesia consisted of Halothane and Nitrous Oxide in all cases. Approximately 2.5 cc of heparinized blood and 0.5 cc of serum were obtained immediately prior to and 2 hours following operation. Half of the children underwent assays of neutrophil function including absolute count, random migration, chemotaxis, phagocytosis, and bacterial killing. Serum was examined for opsonization of Staphylococcus aureus using the chemiluminescence method. The remaining children underwent lymphocytic quantitation including absolute count, total T cells, total B cells, T-helper cells, T-suppressor cells, and T-helper/suppressor ratio. Absolute neutrophil count increased 2.4 times preoperative values (P less than 0.01). There were, however, no significant alterations in neutrophil functional capabilities. Similarly, there was no alteration in serum opsonic capacity. There was a significant decrease in absolute lymphocyte count (6560-4013, P less than 0.01) postoperatively, and T cells, T-helper, T-suppressor, and B cells were all significantly affected (P less than 0.01 to 0.02). There was no change in the T-helper/suppressor ratio.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunotoxicology of brown recluse spider (Loxosceles reclusa) venom.

A purified toxic protein from Loxosceles reclusa venom was assayed for its in vitro effects on the human immunological and blood clotting systems. The toxin caused inhibition of neutrophil chemotaxis, depletion of serum hemolytic complement, prolongation of the activated partial thromboplastin time and depletion of clotting factors XII, XI, IX and VIII by an average of 44% in human plasma. The prothrombin time of human plasma was also prolonged by 1.5-2.0 sec. No effect of the purified toxin was observed on microbicidal ability of neutrophils, the release of enzymes from neutrophils or the adherence of neutrophils to glass beads.

Intravenous immune globulin for hypogammaglobulinemia: A comparison of opsonizing capacity in recipient sera

Abstract Twelve severely hypogammaglobulinemic patients received infusions of alkylated immune globulin and two other native nonalkylated products. Administration was separated by an interval of 3 weeks. Serum was obtained prior to and at 24 hr and 3 weeks after each infusion for measurement of total IgG, specific and opsonizing antibodies. The latter was accomplished against Streptococcus pneumoniae types 5, 12F and 14 and zymosan using chemiluminescence methodology. Changes in total IgG concentrations were comparable for the three products. Prior to enrollment, IgG levels averaged 115 ± 72 mg/dl, increasing to 779 ± 399 at 24 hr postinfusion, and were 337 ± 200 after 3 weeks. No differences among the products were seen in their ability to produce antibodies against Herpes simplex virus types 1 and 2, rubella, toxoplasma, cytomegalo-virus, or tetanus. However, differences in opsonizing antibody were observed between alkylated and native IgG preparations. Peak chemiluminescence responses of neutrophils following opsonization of S. pneumoniae with native immune globulin were significantly higher than with alkylated IgG, indicating greater functional capacity. These studies suggest that native immune serum globulin provides a greater potential for augmenting host defecse mechanisms against pneumococcal infection in hypogammaglobulinemic patients.

Age-depedent variation of lymphocyte function in the postoperative child*

Circulating lymphocyte profiles and reactivity normallyvary with age. Operation results in depression of both lymphocyte counts and blastogenesis but the relationship of age to these alterations has not been previously evaluated in the pediatric surgical patient. This report analyzes the relationship of age to lymphocyte alteration in the postoperative child. Thirty-five healthy children (age range 1 mo to 12 yr), admitted for elective herniorrhaphy, underwent perioperative lymphocyte assay. Anesthesia consisted of halothane and NO 2 , and operative time averaged 54 minutes. Three milliliters of heparinized whole blood was obtained at induction of anesthesia and at 2 hours postoperatively. Lymphocytes were separated by Histopaque gradient and assayed for absolute count, total T-cells, and total B-cells. Lymphocyte reactivity was determined by 3 H-Thymidine incorporation during incubation with the mitogens phytohemagglutinin (PHA), pokeweed (PWM), and/or concanavalin A (Con A) and results expressed as the logarithm of scintillation counts per minute. Differences in preoperative and postoperative values were analyzed for significance by paired T-test. The same differences were evaluated for relationship to age by regression analysis. Operation resulted in significant decreases in all lymphocyte counts (absolute, T- and B-cells), and the operative-induced alteration in both absoluto and total B-cell counts were significantly correlated with age ( P P P

Functional bacterial opsonic activity of human amniotic fluid

There are some data to suggest that amniotic fluid protects the fetus from invasion by pathogenic bacteria. To examine methods by which amniotic fluid may offer such protection, quantitative antibody, complement activity, and functional opsonic capacity were measured. Immunoglobulins were measured by laser nephelometry and total hemolytic complement was determined by radial diffusion; results suggested activity adequate for bactericidal capacity. The chemiluminescence assay was used to quantitate the functional interaction between polymorphonuclear leukocytes and E. coli, group B streptococci (GBS), or zymosan particles preopsonized with amniotic fluid obtained at different stages of gestation. Results were compared to those for normal serum. Data were analyzed by evaluation of the initial slope, area under the curve, and peak chemiluminescence response. Opsonic activity of amniotic fluid for E. coli and GBS was demonstrated, with E. coli showing greater reactivity (maximum = 15,000 to 25,000 cpm) than GBS (10,000 to 20,000 cpm). Specific, as well as nonspecific, opsonic activity was demonstrated by absorption of amniotic fluid with killed bacteria. Concentration of amniotic fluid did not result in an increase in chemiluminescent activity, which demonstrates that optimal opsonic activity already exists. The classical and alternate pathways of complement were assessed for E. coli and GBS. Preterm amniotic fluid did not differ in response from that of amniotic fluid obtained from term pregnancies. This study demonstrates that amniotic fluid can provide the fetus with protection from bacterial pathogens and delineates mechanisms for such protection.

INTRAVENOUS IMMUNE SERUM GLOBULIN FOR HYPOGAMMAGLOBULINEMIA: A COMPARISON OF OPSONIZING CAPACITY IN RECIPIENT SERA

Twelve severely hypogammaglobulinemic patients received infusions of alkylated immune globulin and two other non-alkylated products. Administration was separated by an interval of 3 weeks. Serum was obtained prior to and at 24 hours and 3 weeks after each infusion for measurement of total IgG, specific and opsonizing antibodies. The latter was accomplished against S. pneumoniae types 5, 12F and 14 and zymosan using chemiluminescence methodology.Changes in total IgG concentrations were comparable for the three products. Prior to enrollment, IgG levels averaged 115±72 mg/dl, increasing to 779±399 at 24 hours postinfusion, and were 337±200 after 3 weeks. No differences among the products was seen in their ability to produce antibodies against Herpes simplex virus types 1 and 2, rubella, toxoplasma, cytomegalovirus or tetanus. However, differences in opsonizing antibody were observed between alkylated and native IgG preparations. Peak chemiluminescence responses of neutrophils following opsonization of S. pneumoniae with native immune globulin were significantly higher than with alkylated IgG, indicating greater functional capacity. These studies suggest that native immune serum globulin provides a greater potential for augmenting host defense mechanisms against pneumococcal infection in hypogammaglobulinemic patients.

SURGICALLY INDUCED IMMUNOLOGIC ALTERATIONS IN PEDIATRIC PATIENTS

Surgery is generally believed to be an immunosuppressant, based primarily on studies of adult compromised patients undergoing major operations. Little information is available regarding the young pediatric surgical patient.Eight infants requiring cardiopulmonary bypass (CPB) for cardiac surgery and 50 (mean age: 20 months) undergoing herniorrhapy were evaluated with an extensive survey of immune function. CPB patients had peripheral blood drawn before surgery, post-sternotomy pre-bypass, post-bypass, and 1, 24, and 48 hours post-surgery. Both groups exhibited an increase in absolute neutrophil counts but no change in neutrophil random migration, chemotaxis or phagocytic capacity. However, bacterial killing was reduced in the cardiac surgery group during and after CPB (p=.005). Opsonizing antibody and complement levels remained unchanged in both groups. There was a significant decrease in absolute lymphocyte count post-operatively with T and B cells, T-helper and T-suppressor cells equally effected (p<0.02). There was no change in the T-helper/suppressor ratio. The subtle changes of neutrophil function seen in CPB patients are unlikely to play a major role in postoperative infection. Lymphocyte changes suggest redistribution in response to the stress of surgery.

1276 FUNCTIONAL BACTERIAL OPSONIC ACTIVITY OF HUMAN AMNIOTIC FLUID

There is some data to suggest that amniotic fluid (AF) protects the fetus from invasion by pathogenic bacteria. To examine methods by which AF may offer such protection, quantitative antibody, complement activity, and functional opsonic capacity were measured. Immunoglobulins were measured by laser nephelometry and total hemolytic complement by radial diffusion; results suggested activity adequate for bactericidal capacity.The chemiluminescence assay was used to quantitate the functional interaction between PMNs and E. coli, group B strep (GBS) or zymosan particles preopsonized with amniotic fluid obtained at different stages of gestation. Results were compared to those for normal serum. Data were analyzed by evaluation of the initial slope, area under the curve, and peak chemiluminescence response.Opsonic activity of amniotic fluid for E. coli and GBS was demonstrated with E. coli showing greater reactivity(maximum cpm= 15-25,000) than GBS(10-20,000 cpm). Specific, as well as nonspecific, opsonic activity was demonstrated by absorption of AF with killed bacteria. Concentration of amniotic fluid did not result in an increase in chemiluminescent activity demonstrating that optimal opsonic activity already exists. The classical and alternate pathways of complement were assessed for E. coli and GBS. Amniotic fluids preterm did not differ from term pregnancies. This study demonstrates that amniotic fluid can provide the fetus with protection from bacterial pathogens and delineate mechanisms for such protection.