Inhibition of antibody-complement-mediated killing of tumor cells by hormones.

Line 1, a chemically induced guinea pig hepatoma, is susceptible to killing by anti-Forssman immunoglobulin M antibody and guinea pig complement. When these tumor cells are pretreated at 37 degrees with 10(-4) to 10(-11) M concentrations of the polypeptide hormone insulin, with the catecholamine L-epinephrine-HCl, or with the glucocorticoid steroids hydrocortisone sodium succinate or prednisolone sodium succinate, the cells show a marked reduction in their suseptibility to killing by antibody and guinea pig complement; pretreatment at 0 degrees is ineffective. Similar results were obtained with another antigenically distinct guinea pig hepatoma (line 10) when tested with anti-Forssman immuno-globulin M or specific antitumor antibodies and human complement. The ability of the hormones to render the cells resistant is dependent on time, temperature, and hormone concentration. The effect of hormone treatment is maximal between 30 and 60 min and is reversible within 4 hr even in the continued presence of hormone. Treatment of line 1 cells with up to 10,000-fold greater concentrations of the less biologically active or inactive analogs, DL-epinephrine, beta-estradiol, testosterone, or proinsulin has no effect on the susceptibility of the cells to killing by antibody and guinea pig complement. The effect of hormone treatment is not due to a direct inactivation of bound or fluid-phase complement components by the hormones or to a decrease in the ability of the cells to bind complement-fixing antibody.

PIP: Various aspects of hormone treatment of tumor cells are reported; it is shown that following treatment with certain hormones, the cells are less susceptible to killing by antibody and complement. The diethylnitrosamine-induced guinea pig hepatoma, designated Line 1, is susceptible to killing by anti-Forssman immunoglobulin M (IgM) antibody and guinea pig complement (GPC) but not by specific antitumor antibody and GPC. The antigenetically distinct Line 10 hepatoma, when sensitized with either antibody, is susceptible to killing by human complement (HUC) but not by GPC. Strain 2 of Servall-Wright male guinea pigs were used. 2 antigenetically distinct diethylnitrosamine-induced hepatic tumors (ascites form), Lines 1 and 10, passed in Strain 2 guinea pigs, were collected and suspended in RPMI 1640-20% FCS. Toxicity assays were performed in VBS-gel. The hormones used were hydrocortisone sodium succinate, prednisolone sodium succinate, NSC9151, bovine insulin, L-epinephrine methyl ether HC1, DL-epinephrine, beta-estradiol, testosterone, pork insulin, chicken insulin, pork proinsulin, pork DAA insulin, and the A and B chains of pork insulin. Tumor cells were cultured in 10-ml volumes of RPMI 1640-20% FCS in plastic Petri dishes. After incubation, cell cultures were washed 5 times in VBS-gel and tested for their susceptibility to killing by antibody and complement. Rabbit antiserum to sheep Forssman antigen was prepared and stored at -20 degrees until used. Tumor specific rabbit Antilines 1 and 10 antisera were prepared and similarly stored. Results of tests show that Line 1 tumor cells incubated in a medium containing the polypeptide hormone, insulin, the catecholamine, L-epinephrine HCl, or the glucocorticoid steroids, hydrocortisone sodium succinate, or prednisolone sodium succinate were rendered resistant to killing byanti-Forssman IgM antibody and GPC. This effect was dependent on hormone concentration, temperature, and time. Effects were reversible. Similar results were obtianed with Line 10 cells under attack by specific antitumor and HUC or anti-Forssman antibodies. Less physiologically active analogs of the hormones did not have this effect. Tumor cells showed maximum resistance within 30-60 minutes of exposure to the hormones and reverted to the sensitive state within 4 hours. Resistance of the cells to killing was observed at 37 degrees but not at 0 degrees. It is concluded that the effect of hormone treatment was not due to a direct inactivation of bound or fluid-phase complement components by the hormones or to a decrease in the ability of the cells to bind complement-fixing antibody.

The uniqueness of human milk. Host resistance to infection.

PIP: Field observations are reported which support an assumption that human breast milk acts benefically through induced host resistance to infection. Though the mechanisms of this induced host resistance to infection are not clearly understood, this article suggests the following factors as important: specific antibodies to infectious agents, influences stimulating or inhibiting certain intestinal microorganisms, and nonspecific antimicrobial factors. Immunoglobulins are present in human milk, with IgA representing the most abundant; the role of complement and immunoglobulin in induction of resistance to infection is well-known. Since all immunoglobulins have antibody activity, it is conceivable that immunoglobulins provide the material for antibodies to various etiological agents; for example, serum IgA contains all types of antibodies; serum IgG has a wide variety of antibodies to viruses, rickettsiae, protozoa, H antigens of Salmonella, and bacterial antitoxins and incomplete Rh antibodies; and the IgM fraction contains antibodies to O antigens of Enterobacteriacae, Rh agglutinins, and syphilis reagins. Another aspect of human milk as an inducer of host resistance is the bifidus factor which promotes development of characteristic microflora; since the bifidobacteria metabolize a variety of sugars, producing large amounts of acetic and lactic acids and trace amounts of formic and succinic acids, these organisms affect pH and, hence, certain resistance factors. Other factors present in milk include lysozyme (muramidase), complement, interferon, and immune cells, all of which promote host resistance to infection.^ieng

Proceedings: An investigation into the immunosuppressive properties of oestrogen.

PIP: An investigation into the immunosuppressive properties of estrogen is reported. Normal and thymectomized mature male rats were given daily intradermal injections of .2 ml arachis oil containing 200 mcg estradiol-17beta/ml for 14 days. Lymphocytes from healthy men were incubated with the rat sera and lymphocyte subpopulations were identified by 3 surface markers: 1) cells forming rosettes with sheep erythrocytes (E-rosettes) as a measure of T lymphocytes, 2) lymphocytes with surface immunoglobulin identified by direct immunofluorescence (IgF), and 3) lymphocytes with receptors for C3 observed by a rosette technique employing sheep eryhrocytes treated with rabbit hemolytic serum and human complement (EAC-rosettes). Comparison of the effects of incubating cells with normal or treated rat serum revealed that E-rosettes dropped from 67.4 to 49% and that lymphocytes lacking all markers rose from 2.4 to 17.7%, respectively. The number of cells forming EAC-rosettes, bearing IgF or having both these markers did not change, however, it was apparent that a greater proportion of lymphocytes expressed both markers after incubation with estrogen-treated rat serum (25.7% compared with 12.1%). It appears that estrogen caused both decreased T lymphocyte response and enhanced B lymphocyte activity resulting in increased expression of related markers. None of these phenomena was apparent when lymphocytes were cultured with sera from estrogen-treated thymectomized rats, suggesting that estrogen affects the thymus such that a factor appears in the blood capable of causing changes in the immune response.^ieng

Humoral aspects of anti-HIV immune responses in Zairians with AIDS: lower antigenemia does not correlate with immune complex levels.

Serological patterns of anti-HIV immune responses of 150 HIV-infected (65 asymptomatic, 19 ARC, 66 AIDS) and 150 HIV-negative healthy Zairians were studied to determine the clinical significance of p24 antigen, and anti-p24 antibody, particularly in relation to p24 relative binding capacity (RBC) and circulating immune complexes (CICs). Levels of p24 antigen, anti-p24 antibody titers, and p24 RBC were evaluated by means of enzyme-linked immunosorbent assay (ELISA). Circulating immune complexes were measured by C1q-binding assay. Human immunodeficiency virus CICs were detected by polyethylene glycol (PEG) precipitation followed by 6 M guanidinium lysis, ELISA, Western blot, or radioimmunoprecipitation of the lysed precipitates. Immunoglobulin levels for IgG, IgM, IgA, and beta 2-microglobulin (beta 2-M) were quantified in all study participants by laser nephelometry and ELISA. All immunoglobulin levels were significantly elevated among HIV-positive vs. HIV-negative individuals. Immunoglobulin levels correlated well with disease progression among infected patients. Similarly, beta 2-M levels were significantly higher in HIV-positive vs. HIV-negative individuals and correlated well with progression to AIDS. Free p24 antigen in serum was detected in 1.33% of all patients. However, p24 reactivity increased to 6% (9 of 150 cases) after PEG precipitation and CIC dissociation. There was a good correlation between p24 reactivity and disease development. High titers of anti-p24 antibody (> 44,100) occurred in at least 80% of all patients, and did not correlate with disease stage. Similarly, more than 60% of patients had high levels of p24 RBC.(ABSTRACT TRUNCATED AT 250 WORDS)

Assessment of different sources of variation in the antibody responses to specific malaria antigens in children in Papua New Guinea.

BACKGROUND: A potential problem for malaria vaccine development and testing is between-host variation in antibody responses to specific malaria antigens. Previous work in adults in an area highly endemic for Plasmodium falciparum in Papua New Guinea found that genetic regulation partly explained heterogeneity in responsiveness. We have now assessed the relative contributions of environmental and genetic factors in total IgG responses to specific malaria antigens in children, and quantified temporal variation within individuals of total IgG responses. METHODS: Total IgG responses against schizont extract, merozoite surface protein-1, merozoite surface protein-2, ring-infected erythrocyte surface antigen, and SPf66 were measured by ELISA. Variance component analysis was used to estimate the variation explained by genetic and environmental factors in these antibody responses. Intra- and inter-class correlations of antibody responses within relative pairs were estimated. We adjusted for age, P. falciparum density, sex and village differences either within or prior to the analysis. RESULTS: For all malaria antigens, temporal variation in the total IgG response was the predominant source of variation. There was substantial familial aggregation of all IgG responses, but it remained unclear how much this clustering was attributable to genetic factors and how much to a common environment in the household. The remaining variance, which could not be explained by either of the above, was very small for most of the antigens. CONCLUSIONS: Temporal variation and clustering of immune responses to specific malaria antigens need to be taken into account when planning, conducting and interpreting immuno-epidemiological and vaccine studies.

Immunological aspects of human milk.

PIP: The general decline in breastfeeding in most parts of the world has become a major health concern in view of the many physiological, biochemical, psychological, cultural, and economic benefits of breastfeeding for a few months after birth. A review of the immunologic aspect of breastfeeding shows that a spectrum of interacting specific and nonspecific antimicrobial resistance factors found in the human milk (e.g., physicochemical properties, immunoglobulins, iron-binding protein, interferon, polymorphonuclear leukocytes, and macrophages) are responsible for the lowered incidence of infections, particularly gastrointestinal and respiratory infections, allergic disorders (e.g., infantile eczema), necrotizing enterocolitis, and sudden infant death syndrome among breastfed babies. Possible disadvantages of breastfeeding are 1) poliovirus antibodies in human milk may interfere with live attenuated poliovirus vaccine; and 2) possible infection of breastfed infant with microorganisms found in human milk (e.g., Hepatitis B surface antigen and rubella virus). In view of the significant advantages of breastfeeding for the infant, breastfeeding all newborns during the 1st year of life is recommended.^ieng

Puerperal sterilization.

PIP: This letter is in regard to a previous article by Scott and Guy questioning the need for Rh immunoglobulin in patients having puerperal sterilization. Previous recommendations have been based on the assumption that exposure to Rho (D) antigen only put the patient at risk of developing anti-D antibodies. Recently it has been shown that development of antibodies to each blood group is not an independent process but depends on a mechanism set off by Rho(D) antigen. Of 200 Rh-negative women who developed anti-D antibodies, 36% also had other red cell antibodies. Rh-negative isoimmunized patients who had been transfused with Rh-positive blood had a benign clinical course, but they were not protected from other antigens. However, the risk is considered small. In a survey of 5437 facilities, it was found that the risk of Rh incompatible transfusions was greatest among facilities performing the fewest transfusions. Rh negative blood has not always been available. Anti-Rh prophylaxis has prevented formation of antibodies after incompatible transufsions.^ieng

Rh immunoglobulin utlization after spontaneous and induced abortion.

To monitor the utilization of Rh immunoglobulin (RhIG), we reviewed the charts of 389 spontaneous and 1350 induced abortion patients treated in 1975 at a metropolitan hospital. The rate of ascertainment of Rh type was significantly higher for induced (99.6%) than for spontaneous abortion patients (95.1%) (P less than 0.001). Utilization of Rh immunoglobulin (RhIG) also was significantly higher for induced (98.9%) than for spontaneous abortion patients (80.6%) (P less than 0.001). Women at risk who did not receive RhIG after spontaneous abortion were mostly young, of low gravidity, and at gestational ages (mean 14.4 weeks) associated with substantial risks of Rh sensitization. Eradication of Rh hemolytic disease requires improvement in the system of identifying and treating patients who need prophylaxis.

PIP: The charts of 389 spontaneous and 1350 induced abortion patients treated at a metropolitan hospital in 1975 were reviewed to assess the extent to which Rh immunoglobulin (RhIG) was utilized. The rate of determination of Rh type was 99.6% for cases of induced abortion and 95.1% for spontaneous abortion (p less than .001). RhIG was utilized in 98.9% of the induced abortion cases and in 80.6% of the cases of spontaneous abortion (p less than .001). Those women who did not receive RhIG after spontaneous abortion were primarily young, of low parity, and at those stages of pregnancy associated with a substantial risk of Rh sensitization (mean 14.4 weeks). The results point out the need for a better system for identifying and treating patients at risk of Rh hemolytic disease.

Sperm antibodies in infertile couples.

PIP: From October 1967 to August 1970 377 infertile couples were tested for sperm antibodies using both sperm-agglutination and sperm-immobilization techniques. 17.2% of the couples had demonstrable sperm antibodies, 11.4% in the wife and 5.8% in the husband. In 20.4% of the cases of primary infertility sperm antibodies were detected. This was 3 times higher than the rate in secondary infertility. Other factors which contributed to infertility in a recent series of 150 cases were low cervical mucus pH, poor liquefacation of the sperm semen sample, and poor sperm penetration of the cervical mucus.^ieng

Ultrasound as a new method of male contraception.

Twenty male cats were treated once or twice with 1 watt/sq cm of ultrasound for 10 minutes. Each of 24 male dogs received one to three treatments with 1 watt/sq cm for 10 minutes. Another six dogs were treated with 2 watts/sq cm for 15 minutes. Four Cebus apella monkeys were treated with the same dosage as that used for the cats and dogs. A dosage of 1 watt/sq cm for 10 minutes was also applied to four human patients without the use of anesthetics, and no pain or side effects were noted. In all treated animals as well as in human patients the results indicate that ultrasound significantly suppresses spermatogenesis according to the dosage and frequency of treatment, without any effect on Leydig cells or blood testosterone levels.

PIP: Ultrasound was tested as a method of sterilization in cats, dogs, monkeys, and humans, utilizing an ultrasound machine manufactured by Whitewater Electronics specifically for this research. 30 male cats were divided into 3 groups, 2 experimental and 1 control. The treated groups received 1 watt/sq cm for 10 minutes once, or a 2nd time after 48 hours. Testicular biopsies were made 60 days after treatment. 24 male dogs were divided into 3 groups receiving either 1,2, or 3 treatments within a 1-week interval with 1 watt/sq cm for 10 minutes. Sperm counts were made for 5 consecutive weeks preceding ultrasound treatment and semen analyses made weekly. 6 dogs were treated with 2 watts/sq cm for 15 minutes. Antibodies to spermatozoa were determined by the Kibrick method. 4 South African monkeys were treated with 1 watt/sq cm for 10 minutes; testicular biopsies were obtained 1 month before treatment and 60 days after treatment. 1 watt/sq cm for 10 minutes. Testicular biopsies were obtained 3 weeks before treatment and orchiectomies were performed 14 -17 days after treatment. In all treated animals and human patients, ultrasound significantly suppresses spermatogenesis accoridng to the dosage and frequency of treatment, without any effect on Leydig cells or blood testosterone levels.

An evaluation of immunologic factors of infertility.

PIP: This paper reviews earlier studies identifying 3 possible immunologic causes of infertility: autoimmunity, ABO blood group incompatibility, and isoagglutination, and presents results of recent studies, including an analysis of the incidence of isoagglutinations, autoagglutinations, and ABO incompatibility in a group of couples with unexplained primary or secondary infertility and couples with an organic cause of infertility; incidence of antibodies in postpartum women, women in early pregnancy, unmarried women, and prostitutes; the relationship between ABO incompatibility and/or sperm-agglutinating antibodies to postcoital testing of cervical mucus; results of condom therapy in couples with positive isoagglutinins; a comparison of hemagglutination test with the microagglutination test; and preliminary studies of the immunoglobulin responsible for a positive microagglutination reaction. Results obtained from 292 women and 176 men indicated that in the 64 couples with primary unexplained infertility, 24 wives had isoagglutinins and 5 of the husbands had autoagglutinins. Among 32 couples with secondary unexplained infertility, 16 wives had isoagglutinins and only 1 husband had autogglutinins. 50 couples with organic causes of infertility and 25 unmarried women had a 20% incidence of isoagglutinins. Positive serologic reactions were found in 35 of 48 prostitutes. 15 of 44 women in the 1st trimester of pregnancy had isoagglutinins to at least 1 of 3-5 donor semen specimens, but serums of 7 of the women gave negative reactions when tested against the husband's ejaculate. The 2 women in the group who aborted had negative antibody titers. Only 1 of 29 serums from postpartum women gave positive agglutination reaction. Among 68 couples, 19 subjects had positive microagglutination test reactions but only 7 had positive hemagglutination reactions. No specific relationship was found between ABO incompatibility and the results of postcoital testing of cervical mucus in 58 women tested or positive circulating issoagglutinins in 62 women tested. 27 of 68 couples with primary unexplained infertility, 18 of 33 with secondary unexplained infertility, and 13 of 46 with organic causes of infertility had ABO incompatiblity. Autoagglutination, unless massive, was not incompatible with pregnancy. Circulating antisperm antibodies fell to undetectable levels after 2-12 months of condom therapy. Careful timing of ovulation is essential because of the increase in antibodies after reexposure. Results of condom therapy in terms of pregnancies were disappointing.^ieng

The clinical relevance of classes of immunoglobulins on spermatozoa from infertile and vasovasostomized males.

Following reversal of vasectomy, conceptions occur even when antisperm antibodies are present in the seminal plasma, but this is most unusual in men with similar titers of such antibodies who are spontaneously infertile. To clarify the differences between antisperm antibodies occurring in infertile men and those associated with vasectomy reversal, we have studied 23 spontaneously infertile men and 22 men who underwent vasectomy reversal, all of whom had antisperm antibodies detected in seminal plasma by the same tray agglutination test. The class of antibody on spermatozoa was defined by a double-antibody technique using diluted rabbit anti-human IgG, IgM, or IgA or secretory component, followed, after washing, by 125I-labeled donkey anti-rabbit Ig. The results have shown that similar amounts of IgG and IgM were present on the spermatozoa, but infertile men had significantly more IgA and especially more secretory component than men who underwent vasectomy reversal. This was associated with significantly greater impairment of penetration of cervical mucus in the former group. It appears that the type of antibody on the spermatozoa may vary according to the stimulus for its production.

PIP: To clarify the differences between antisperm antibodies occurring in spontaneously infertile men and in men who have undergone reversal of vasectomy, classes of immunoglobulins were compared in 23 men from the former group and 22 men from the latter group. Antisperm antibodies were detected in seminal plasma by the same tray agglutination test in all subjects. The class of antibody on spermatozoa was defined by a double-antibody technique that used diluted rabbit antihuman IgG, IgA, or secretory component, followed, after washing, by 125I-labled donkey antirabbit Ig. Although similar amounts of IgG amd IgM were present on the spermatozoa in both groups, infertile men had significantly more IgA and especially more secretory component than men who underwent vasectomy reversal. This was associated with significantly greater impairment of penetration of cervical mucus in the former group. 8 of the 20 vasovasectomized men with antisperm antibodies in their seminal plasma produced pregnancies, whereas no pregnancies have been observed in 63 spontaneously infertile untreated men with seminal plasma antibodies. Spermatozoa from 86% of vasovasectomized men penetrated the cervical mucus, while spermatozoa from only 27% of spontaneously infertile men were able to do so. It is concluded that the type of antibody on the spermatozoa may vary according to the stimulus for its production.

Contraception and cervical colonization with mycoplasmas and infection with cytomegalovirus.

The relationship of cervical colonization of genital mycoplasmas and infection with cytomegalovirus (CMV) was studied in 66 intrauterine device (IUD) users as contrasted to 60 patients using oral contraception and 50 patients using neither an IUD nor oral contraception. No significant increase in colonization or genital mycoplasmas or infection with CMV was noted in IUD users. No CMV was isolated from users of the cooper-t IUD. It is unlikely that genital mycoplasmas are related to either the increased immunoglobulin levels seen in patients using the IUD or the antifertility effects of the IUD.

PIP: An ivestigation based on the hypothesis that the IUD might induce an endometritis related to a nonbacterial infection was undertaken in 66 IUD users and contrasted to 60 oral contraceptive (OC) users and 50 nonusers of OCs and IUDs. The relationship of cervical colonization of genital mycoplasmas and infection with cytomegalovirus (CMV) was studied yielding the following information: 1) Genital mycoplasma colonization among the groups studied differed insignificantly; 2) An insignificant increase in colonization of genital mycoplasmas or infection with CMV was noted in IUD users; 3) There were no CMV isolates from the group using the Copper T IUD; 4) CMV infection did not appear to be significantly increased by IUD presence; and 5) The overall rate of 5.2% of CMV infection was comparable with that of other studies. Results indicated the unlikelyhood that genital mycoplasmas are related to either the increased immunoglobulin levels noted in patients using the IUD or to the IUD's antifertility effects.

A new look at antifertility vaccines.

PIP: This article reviews new advances in biochemistry, biotechnology, and immunology relevant to antifertility vaccine development and evaluates the current status and future prospects of contraceptive vaccines and other immunologic approaches to fertility regulation. Contraceptive vaccine candidates include human chorionic gonadotropin, human luteinizing hormone and luteinizing hormone releasing hormone, and reproductive steroid hormones. Sperm enzymes are attractive for a contraceptive vaccine; among the sperm antigens studied are antibodies to hyaluronidase, acrosin, and lactate dehydrogenase-C4. Several laboratories have developed monoclonal antibodies to a variety of sperm antigens and are using them to identify and characterize new sperm proteins and their roles in fertility. Considerable progress has been made toward biochemical characterization of unique glycoproteins constituting the zona pellucida. Zona pellucida antigens are good candidates because antizona antibodies may block both fertilization and implantation, and low amounts of antibody would be sufficient because of the small number of mature eggs with zona present at any time. Studies are underway to identify human embryonic antigens through examination of the protein profile of human teratocarcinoma cell lines at various stages of differentiation and through analysis of antibodies in human pregnancy and infertility sera. Placental and extraembryonic membranes produce several tissue-specific antigens that have been considered for antifertility vaccines, but concern that they could produce late or incomplete abortion has prevented their serioud consideration. Because of possibly serious systemic side effects, presence of the blood-testis barrier, and large number of sperm produced daily, it is unlikely that sperm vaccines can be safely administered to men. Nautural protective mechanisms will probably render some immunocontraceptive approaches ineffective. The possibility of serious pathogenic side effects of contraceptive vaccines demands vaccines demands a cautious approach to their development.^ieng

Infection with Pneumocystis carinii is prevalent in healthy Gambian children.

Pneumocystis pneumonia is rarely identified in the many immunosuppressed individuals with acquired immune deficiency syndrome (AIDS) and malnutrition in Africa. To test whether infection with Pneumocystis carinii occurs in the continent we conducted a comparative serological study, measuring by enzyme-linked immunosorbent assay antibodies to the parasite in 150 healthy young individuals from both Britain and the Gambian savanna. The prevalence of significant titres of antibody to P. carinii steadily increased with age and included more than 70% of both populations by 8 years of age. Infection with P. carinii is, therefore, common in the Gambia. Thus opportunistic pneumocystis pneumonia may be an important but largely unrecognized disease in the continent, though its impact is probably diminished by the prevalence of fatal tuberculous infection, particularly in the AIDS population.

PIP: The prevalence of sero-positivity toward Pneumocystis carinii parasite antigens was determined in 150 British subjects aged 3 months-40years, and in 150 Gambians of the same age age range. The Gambians were from Katabas villages north of the Gambian river. The sera were tested with an ELISA assay developed from parasites and cysts isolated from rats, which were previously shown to share many antigens with organisms from humans. IgG titers reactive with P. carinii increased with age in the British and Gambian subjects, with essentially no positives at ages 3-6 months, to a peak of 70% positive in both groups by age 8 years. British children and parents were questioned about history of pneumonia illness, with negative findings. There are very few reports of P. carinii infections, judging from sputum and bronchoscopic lavage, in African AIDS patients. 2 cases of Cryptococcus have been reported, but tuberculosis is a common and rapidly lethal infection in African AIDS patients.

Severe vitamin A deficiency in a rural village in the Hararge region of Ethiopia.

A total of 240 children were examined for vitamin A deficiency in a village in Hararge region of Ethiopia. Night blindness, Bitot's spots, corneal xerosis, corneal ulceration and corneal scars were observed in 69, 16, 2, 15 and 14 children, respectively, based on the most severe eye signs. Blood was collected from 76 children with eye signs and 9 other children selected at random. The concentration of retinol (vitamin A) in serum was < 0.35 mumol/l in 30.2% of children and the median serum retinol-binding protein, iron, transferrin saturation and ferritin levels were low while the parameters of iodine status, total triiodothyronine, total thyroxine and thyrotropin, were all within the normal range. Levels of IgG and IgM were elevated in 78.8% and 82.4% children, respectively, while C-reactive protein levels were elevated in 42.4% of children. There was a higher prevalence of wasting (33%) than stunting (10%) with an additional 8% of children being both stunted and wasted. In the 2 years prior to the study, there were 74 deaths of which 17 were reported to be associated with ruptured corneas. The community had been dependent on relief food aid for the previous 6 years.

PIP: In late 1988 in Ethiopia, a physician and a nutritionist examined 240 registered children in the village of Melkaye (Farmers' Association No. 34) in Darolobo District of Habro Province for vitamin A deficiency. These children had been dependent on food aid because of consistent drought and crop failures since, at least, 1982. The food aid, which was the main food source, included food deficient in vitamin A and beta-carotene: wheat flour, vegetable oil, butter-oil, and beans. 53.2% of the boys and 43.1% of the girls had at least 1 sign of vitamin A deficiency, especially night blindness, an early manifestation of vitamin A deficiency. 28.8% of all children had night blindness without signs of xerophthalmia compared with the WHO cut-off point of 1%. (The cut-off point is used to determine the public health significance of vitamin A deficiency.) 6.7% had Bitot's spots compared with a cut-off point of 0.5%. 7.1% had corneal xerosis/ulceration compared with a cut-off point of .01%. 5.8% had corneal scars compared with a cut-off point of .05%. 30.2% had a serum retinol level less than .35 mcmol/l compared with a cut-off point of 5%. 17 of 70 children (24.3%) who had died in the last 2 years had ruptured or damaged eye(s). The median levels of serum retinol-binding protein, iron, transferrin saturation, and ferritin were lower than normal levels. On the other hand, parameters of iodine status, total triiodothyronine, and total thyroxine and thyrotropin were all normal. 78.8% and 82.4% of the children experienced high levels of IgG and IgM, respectively. 42.4% had high C-reactive protein levels. Wasting was more common than stunting (33% vs. 10%). 8% suffered from both stunting and wasting. The severity of xerophthalmia was perhaps the most severe ever recorded and prompted health workers to distribute vitamin A capsules to all children in Melkaye and nearby villages.

Induction of aryl hydrocarbon hydroxylase by lymphocytes from women taking oral contraceptives.

The peripheral blood lymphocytes (PBL) from two female subjects were assayed for AHH induction 40 days prior to and 30 days during ingestion of progesterone and estrogen analogues as oral contraceptives. Three habitual users of oral contraceptives were also studied. No in vitro inhibition of AHH induction was observed as a consequence of the use of these hormone analogues. Values obtained for enzyme activity suggest a slight increase in AHH induction resulting from the use of oral contraceptives. Further studies with larger numbers of subjects are required before the apparent increase in enzyme inducibility can be considered significant.

PIP: 2 healthy female volunteers were tested 3 times weekly to determine any effects of ingestion of a combined oral contraceptive (OC) agent on the inducibility of aryl hydrocarbon hydroxylase (AHH), an enzyme known to be readily synthesized by human lymphocytes in the presence of carcinogens. These 2 subjects were assayed 40 days before and 30 days during ingestion. 3 habitual users of OCs were also studied. In the absence of OCs, the 2 highest values occurring at or immediately following the approximate time of ovulation. In the presence of a lower dose OC (.3 mg ethinyl estradiol and .3 norgestrel), the daily levels of AHH inducibility followed a cyclical pattern very similar to that found in the absence of OCs, with a peak at Day 17. On higher dosage (.5 mg ethinyl estradiol and .5 mg norgestrel), a greater degree of variability was noticed, but the overall inducibility pattern followed the same general trend. Hence, no in vitro inhibition of AHH induction was observed as a consequence of using hormone analogs; but the values, shown graphically, obtained for the enzyme activity suggested a slight increase in AHH induction resulting from the OC use. Further, larger studies are called for.

Humoral immunity in oral contraceptive users. I. Plasma immunoglobulin levels.

The worldwide acceptance of steroid-based oral contraception makes it imperative that the effect of these agents on the immune system is understood. Nevertheless, information regarding the effect of steroid-based oral contraception on plasma immunoglobulin (Ig) levels is often conflicting. In this report immunoglobulin levels in the plasma of females taking steroid-based oral contraceptives are measured in a novel manner using an enzyme-linked immunosorbent assay (ELISA). No significant alterations in the levels of IgG or IgM are reported.

PIP: Immunoglobulin IgG and IgM levels in 10 women taking a variety of oral contraceptives containing ethinyl estradiol and levonorgestrel were assayed by the sensitive and specific ELISA method. 10 women matched for age and socioeconomic status served as controls. No significant differences in IgG or IgM levels were observed. Although several reports in the literature, using less sensitive methods such as immunodiffusion and immunoelectrophoresis, also reported no difference, there have been a few conflicting reports of alterations in women taking pills.

Humoral immunity in oral contraceptive users. II. In vitro immunoglobulin production.

The worldwide acceptance of steroid-based oral contraception makes it imperative that the effect of these agents on the immune system is understood. Nevertheless, information regarding the effect of steroid-based oral contraception on humoral immunoregulation is limited. In this report the in vitro production of IgG and IgM is measured following stimulation with either the T-dependent activator pokeweed mitogen (PWM) or the T-independent activator fixed/killed Staphylococcus aureus Cowan I (StaCw). No significant differences are observed between the in vitro IgG or IgM levels following stimulation with PWM or StaCw for females taking steroid-based oral contraceptives and females not taking steroid-based oral contraceptives. We conclude that humoral immunoregulation is unaltered in steroid-based oral contraceptive users.

PIP: Production of IgG and IgM immunoglobulins in vitro by purified monocytes isolated from the blood of women taking oral contraceptives was assayed by the sensitive ELISA method. The subjects were taking pills containing ethinyl estradiol and levonorgestrel. Washed mononuclear cell suspensions containing no more than 5% polymorphonuclear cells were incubated 7 days with either formaldehyde-killed Staphylococcus aureus strain Cowan I or T-cell dependent pokeweed mitogen. No significant differences were observed in immunoglobulin production between pill users and controls.

Immunoglobulin levels in the serum and cervical mucus of tailed copper IUD users.

Immunoglobulin levels were measured by agar gel single radial immunodiffusion in the serum and cervical mucus of 50 tailed copper IUD users, 20 combined oral contraceptive users and 20 women not using contraception (controls). IgG, IgA and IgM levels were significantly higher in the copper IUD users compared to the other two groups. It is not known whether this was related to the small numbers of bacteria found in the uterine cavity of tailed IUD users or to the foreign body reaction of the device.