Screening for gestational diabetes mellitus: a critical review

Bovine haptoglobin, which has been recognized as an acute phase protein following tissue injury and inflammation, was detected as a 33 k + 20 k Dalton fraction in the sera from calves transported by road for 2 days. The sera also possessed suppressive activity on lymphocyte proliferative response to concanavalin A. A significant correlation (r = 0.57, P < 0.05) was observed between haptoglobin concentrations and lymphocyte suppression in the sera. Furthermore, the haptoglobin fraction obtained from acute phase sera exerted dose-dependent suppression on lymphocyte blastogenesis. These circumstantial data suggest the possible involvement of bovine haptoglobin, at least in part, as an immunomodulator in serum suppression of lymphocyte blastogenesis in transported calves.     

Lymphocyte function in experimental african trypanosomiasis: mitogenic effects of trypanosome extracts in vitro

Extracts of Trypanosoma brucei and Trypanosoma congolense were incubated in vitro with nonimmune lymphocytes of mice, rats, guinea pigs, and rabbits in order to test for mitogenic effects or for other characteristics of polyclonal B lymphocyte activators. Trypanosome extracts (TE) were not mitogenic for spleen cells of mice, rats, and guinea pigs in vitro, nor did the parasite extracts alter the mitogenic responses of lymphocytes from these animals to known B- and T-cell mitogens. TE also failed to induce polyclonal antibody synthesis in mouse spleen cell cultures in an in vitro antibody response system, in contrast to the effects of bacterial lipopolysaccharide, a known polyclonal B cell activator. Rabbit spleen cell and peripheral blood lymphocyte cultures, however, were stimulated by TE to undergo blastogenesis in vitro. Incubation of rabbit lymphocytes with phytohemagglutinin (PHA) and TE or anti-rabbit immunoglobulin serum and TE revealed an additive effect only in terms of the TE-plus-PHA culture responses; these findings suggest that a non-PHA responsive lymphocyte population, possibly B lymphocytes, is stimulated by TE in rabbits. The relationship of trypanosome-induced lymphocyte mitogenic stimulation to other immunological dysfunctions occurring in chronic African trypanosomiasis is discussed.     

Exophiala dermatitidis and Sarcinomyces phaeomuriformis: ITS1-sequencing and nutritional physiology

Measurement of the T cell blastogenic response to Candida may be useful in the evaluation of patients with suspected immunodeficiency. The classic blastogenesis assay is based on uptake of [3H]thymidine by peripheral blood lymphocytes stimulated with Candida antigens for 5 days. An alternative approach involves staining peripheral blood lymphocytes with the intracellular fluorescent dye carboxyfluorescein succinimidyl ester (CFSE) and measuring mitotic activity by the successive twofold reductions in fluorescent intensity using flow cytometry (FCM). The two approaches were compared in 16 subjects who demonstrated various proliferative responses to Candida. FCM-derived indices all involved initial gating on CD3+ T cells and included 1) blastic transformation as measured by changes in light scatter, 2) cell division, measured by CFSE fluorescence, and 3) CD69 expression. A good correlation was found between [3H]thymidine uptake and CFSE-derived indices, irrespective of the analysis algorithm used to interpret CFSE division profiles. Furthermore, significant T cell proliferation occurred only in subjects who had had one or more symptomatic episodes of vaginal candidiasis whereas controls with no such history, and patients with chronic vaginal infection, showed minimal proliferation. The increase in proportion of CD69+ T cells in culture also correlated with the blastogenic response to Candida, but less well than mitotic indices. CFSE-derived indices of T cell blastogenesis to Candida are equivalent to [3H]thymidine-based assays and may allow useful laboratory distinction between subjects who have been exposed to and recovered from vaginal Candida infection, who have a strong proliferative response, from those with no exposure or chronic infection who demonstrate a poor response.     

HIV-specific lymphoproliferative responses in asymptomatic HIV-infected individuals

In vitro lymphoproliferative responses to HIV-1 recombinant antigens (gp160, p24, and Rev protein) were studied in 83 patients with asymptomatic HIV-1 infection (CDC groups II and III) and circulating CD4 lymphocyte numbers > 400/mm3. Significant response to at least one of the three antigens was detected in 52.4% of the subjects, but the responses were weak, and concordance of the response to the three antigens was rare, the frequency of individuals responding to each antigen not exceeding 22.4%. Increasing frequencies of response were observed when recall antigens (tetanus toxoid and Candida albicans glycomannoprotein) (65.5%) and anti-CD3 MoAb (76.6%) were used as stimuli. Although a significant association between lymphocyte response to p24, but not gp160, and steadiness of CD4 lymphocyte numbers before the assay was observed, no predictive value for lack of CD4 cell decrease was confirmed for either antigen, and fluctuation of the responses to HIV antigens was seen during subsequent follow up. The panel of T cell assays used could be regarded as appropriate for monitoring both HIV-specific responses and T lymphocyte function during immunotherapy with soluble HIV antigens.     

Longitudinal studies of immune function in cattle experimentally infected with bovine immunodeficiency-like virus and/or bovine leukemia virus

The effects of single or dual infection with bovine immunodeficiency-like virus (BIV) and/or, bovine leukemia virus (BLV) on bovine immune function were examined over a 4 year period. Holstein calves were infected with BIV (four calves), BLV (five calves), BIV and BLV (five calves), or sham inoculated (three calves). Lymphocyte blastogenesis to mitogens, seven tests of neutrophil function, and mononuclear cell subset analysis by flow cytometry (BoCD4, BoCD8, BoCD2, BoWC1, sIgM+, and monocytes) were performed at regular intervals to 49 months post-infection. These data were analyzed for main effects of each virus and interaction as a 2 x 2 factorial. BIV infected cattle had lower neutrophil antibody-dependent cell-mediated cytotoxicity and iodination responses during 2 of the 4 years post-infection (P < 0.05). BIV infection was not associated with any long-term significant changes in lymphocyte blastogenesis to mitogens or changes in mononuclear cell subset numbers in blood. There was a tendency for animals infected with BIV alone to have decreased lymphocyte blastogenic responses to mitogens, but this was not statistically significant. BLV infection caused an increase in total mononuclear cells with no dramatic shift in the relative proportions of the various subsets. Co-infection with BIV and BLV did not consistently cause a different response than either virus did individually. One BIV infected animal died of non-BLV lymphosarcoma 7 months after infection. All other animals had no unusual clinical signs. In summary, infection with BIV caused a significant, temporary decrease in neutrophil function with no consistent statistically significant alteration in lymphocyte blastogenesis or mononuclear cell numbers during the first 4 years after infection. BLV infection caused an increase in lymphocyte numbers, and there appeared to be no synergism between the viruses.     

Lymphocyte transformation in vitro in dermatophytosis

Peripheral blood lymphocytes from 59 patients with dermatophytosis and from nine young healthy women were studied by the lymphocyte transformation test (LT) using mitogens and bacterial as well as fungal antigens. The latter included Candida albicans (CA) and four dermatophyte species, viz. Trichophyton rubrum (TR), Trichophyton mentagrophytes (TM), Epidermophyton floccosum (EF) and Microsporum canis (MF). Most of the patients showed normal transformation in response to mitogens and non-dermatophyte antigens, indicating that they have no functional T-cell deficiency. Dermatophyte antigens act as stimulators in LT. In general, patient lymphocytes responded more strongly to these antigens than lymphocytes from controls. In most patients suffering from TM infections, response to the TM antigen was significantly stronger (p less than 0.05) than that in the other patients, indicating that this antigen preparation shows species specificity. In patients with Trichophyton (TR + TM) infections, response to the corresponding antigens was significantly stronger than that in the other patients, which suggests the existence of genus specificity. Any differences between patients suffering from chronic TR infections and those with acute TR infections were not observed, a finding which is in contrast to those obtained in other studies. However, a few patients with chronic TM infections responded weakly to mitogens and non-dermatophyte antigens. LT in four patients with id-reaction to TM infection was not found to differ from that in the remaining TM patients.     

Color-flow duplex ultrasound assessment of internal thoracic artery graft after coronary bypass

Cell-mediated immune responses to chlamydial and common recall antigens were measured in 26 subjects whose clinical signs of trachoma persisted over 6 months of follow-up and in 21 subjects whose clinical signs resolved spontaneously over the same period. Seven-day lymphocyte proliferative responses to chlamydial but not common recall antigens were significantly greater in subjects whose disease resolved spontaneously. There was, however, no detectable difference between the two groups in gamma interferon levels in supernatants from lymphocyte cultures stimulated with these antigens. These results are consistent with the hypothesis that cell-mediated immune responses play an important role in the clearance of ocular chlamydial infection in humans.     

Selective elimination of the delayed-type hypersensitivity response by extracorporeal thoracic duct filtration

Calves were sensitized to tuberculin and histoplasmin. The delayed-type hypersensitivity skin response to these antigens was produced and the diameter of induration measured. Repeated skin tests prior to filtration demonstrated that the amount of induration produced by these skin tests was closely reproducible. Histoplasmin or tuberculin-coated columns were then introduced into a closed circuit extracorporeal thoracic duct circulation. A significant (P is less than 0.01) reduction or ablation of the delayed-type hypersensitivity response was obtained to the antigen used to coat the column. In contrast, no significant reduction occurred in the skin test response to the other antigen. Repeated skin tests to both antigens after the cessation of filtration showed a gradual rise toward prefiltration levels in the skin test to the filtered antigen. The results of these experiments indicate that a selective population of T lymphocytes can be removed from an in vivo system. The removal of these cells can selectively reduce a delayed-type hypersensitivity skin test response to a particular antigen. These results are consistent with the hypothesis that a type of in vivo selective immunosuppression can be produced by antigen-coated columns when they are placed in an extracorporeal thoracic duct lymph circulation system.     

Effect of pentachlorophenol on immune function

The organochlorine compound, pentachlorophenol, was evaluated for effects on immune system function in male Fisher 344 rats. Pentachlorophenol was prepared in an olive oil vehicle and was administered by oral gavage twice weekly for 28 days at a dose of 2.0 mg/kg per treatment. Exposure to pentachlorophenol increased body weight gains (P=0.024) during the treatment period. Liver (P=0.034) and kidney (P=0.012) body weight ratios were also increased. Pentachlorophenol exposure enhanced T-lymphocyte blastogenesis induced by concanavalin A (Con A)(P=0.0001) and phytohemagglutinin (PHA)(P=0.048) evaluated using stimulation indices. Corresponding B-lymphocyte blastogenesis induced by lipopolysaccharide/dextran (LPS/dex)(P=0.0034) was also enhanced by pentachlorophenol exposure. Pentachlorophenol suppressed the antibody response against sheep red blood cells (SRBCs) by 39% when the response was expressed per viable spleen cell (P=0.006). This suppression was not evident when the response was expressed per spleen (P=0.22), suggesting that a compensatory mechanism or extramedullary splenic hemopoiesis was occurring minimizing the overall impact on humoral immunity. The enhanced B- and T-lymphocyte blastogenesis may also reflect compensatory or hemopoietic activity. Pentachlorophenol exposure had no effect on peritoneal macrophage phagocytosis (P=0.31) or lymphocyte cell surface antigen expression. The observed alterations in lymphocyte blastogenesis and humoral immunity subsequent to pentachlorophenol exposure do not appear to be associated with phagocytosis or lymphocyte cell surface antigen expression.     

Immune response profile in patients with active tuberculosis in a BCG vaccinated area

Tuberculosis patients with pulmonary (N = 95) or lymph node disease (N = 23) were assessed for Th1 responses (PPD skin test and lymphocyte blastogenic and interferon gamma) and Th2 responses (polyclonal and antigen specific IgE). Skin test responses to PPD and lymphocyte proliferative responses to crude mycobacterial antigens (PPD, culture filtrate and sonicate) and recall antigens (tetanus toxoid and streptolysin O) were significantly suppressed (p < 0.001) in patients with pulmonary disease compared to endemic controls. However, mitogen (phytohemagglutinin)-stimulated responses were comparable in patients and controls. Polyclonal and antigen specific (M. tuberculosis culture filtrate) IgE responses which are considered to be surrogate markers for Th2 responses were significantly higher in patients with pulmonary disease compared to healthy endemic controls (Mann Whitney analysis p < 0.01). Patients with lymph node disease showed strong Th1 responses but did not show significant responses for either polyclonal or antigen specific IgE. Thus overall suppression of T cell memory response was observed only in patients with pulmonary disease but not in patients with lymph node disease suggesting that sequestration of antigen in different compartments leads to differential activation of Th1 and Th2 responses. PPD skin test responses were highly positive in endemic controls (47% positive) and household contacts (86% positive). Furthermore, PPD positivity decreased with disease severity. Therefore PPD positivity in a BCG vaccinated TB endemic area cannot be used as a diagnostic marker for active tuberculosis particularly in advanced disease.     

Studies on Pasteurella multocida. III. In vitro assay for cell-mediated immunity

Peripheral blood lymphocytes (PBL) from turkeys immunized against fowl cholera with a bacterin or a live avirulent vaccine (strain CS-148) were cultured in vitro with various antigenic preparations from Pasteurella multocida (strain P-1059). The degree of lymphocyte stimulation (blastogenesis) was quantitated by measurement of the uptake of (3H) thymidine. Higher stimulation indices were obtained with immune lymphocytes rather than nonimmune lymphocytes. Stimulation was specific since PBL from turkeys immunized against P. multocida failed to react with Escherichia coli or Mycoplasma synoviae antigens. These differences were statistically significant as analyzed with the student's t-test. The lymphocyte transformation assay was emphasized as a convenient and useful in vitro indicator of cell-mediated immunity that should help define the role of cell-mediated immunity in P. multocida infections of turkeys.     

Immunologic protection afforded by sunscreens in vitro

Several studies have suggested a lack of correlation between sunscreen sun protection factor and protection of the skin immune system, potentially allowing greater damage to the skin by removing the natural protective erythemal response to sun exposure. Despite this, routine testing of immune protection afforded by sunscreens is not performed by industry. Current laboratory methods for investigating the efficacy of sunscreen protection of epidermal immune function use the induction of contact hypersensitivity or epidermal cell alloantigen presentation. Animal models, cell culture systems, and in vivo human studies are commonly employed, but all these systems have significant drawbacks for use in routine testing. The purpose of this study was to develop an in vitro system for testing the immunologic protection afforded by sunscreens in human skin. Five test sunscreens plus a vehicle control were tested in a "blind" fashion for their in vitro level of immune protection. Creams were applied in a standard manner to human whole skin explants and were irradiated over a range of physiologic doses using an Oriel solar simulator. A mixed epidermal lymphocyte reaction was used to quantify epidermal alloantigen-presenting capacity, in the presence or absence of test cream, for five explants. Results consistently demonstrated that all the test sunscreens protected beyond their designated sun protection factors, whereas the vehicle conferred no protection. The explant-mixed epidermal lymphocyte reaction system gave consistent, reproducible results and may prove useful for the allocation of an immune protection factor to all sunscreens.     

Relation between fat intake and mortality: an ecological analysis in Belgium

To understand the role of the motor cortex in implicit and explicit learning, we studied alpha event-related desynchronization (ERD) while 13 right-handed individuals performed a variation of the serial reaction time task (SRTT). EEG signals were recorded simultaneously from 29 scalp locations and the ERD was computed. During data collection, all subjects developed implicit knowledge, demonstrated by shortening of the response time, and explicit knowledge of the test sequence. The average ERD maps of all 13 subjects demonstrated that during the initial learning, there was a decline in alpha band power that was maximal over the contralateral central region. The ERD reached a transient peak amplitude at a point when the subjects attained full explicit knowledge, and diminished subsequently. The transient peak in ERD was highly significant at C3. These electrophysiologic findings support previous studies which have demonstrated that motor activity changes as behavior changes over the course of learning.     

Effect of dietary restriction on cell-mediated immune responses in cattle infected with Mycobacterium bovis

Nine M. bovis-infected cattle on a diet deficient in both protein and energy for 133 days lost approximately 17% of their original body weight. However, dietary restriction did not result in any significant reduction in skin sensitivity to PPD, in vitro production of IFN-gamma or lymphocyte blastogenesis. The number of circulating BoCD4+ cells and B cells were similar in both the malnourished and the control cattle. However, significantly lower numbers (P < 0.01) of circulating BoCD2+ cells, BoCD8+ cells, WC1+ gamma delta T cells and ACT2+ cells were found in the malnourished cattle. With the exception of inorganic phosphate, the changes in plasma biochemical parameters were unremarkable.     

Instability of tuberculin and Candida skin test reactivity in HIV-infected Ugandans. The Uganda-Case Western Reserve University Research Collaboration

Anergy testing has been used as an adjunct to tuberculin testing for assessing M. tuberculosis (MTB) infection and indications for isoniazid preventive therapy in HIV-infected persons. We examined factors associated with the stability of skin test responses to purified protein derivative (PPD) and candida antigens in a cohort of HIV-infected adults followed prospectively in a tuberculosis preventive therapy trial in Uganda. PPD-positive and anergic subjects in the placebo arms of the preventive therapy study underwent repeat skin testing and immunologic testing including measurement of MTB culture filtrate (CF)-stimulated interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) levels in whole-blood culture supernatants. Anergy was present in 27% of 4,058 HIV-infected subjects screened for the tuberculosis preventive therapy trial compared with 10% of 682 HIV-non-infected persons. On follow-up testing of enrolled subjects, 42% of 139 initially anergic subjects were no longer anergic; two thirds of these had PPD reactions >= 5 mm. Stability of anergy was associated with intercurrent opportunistic infections and AIDS-associated dermatitis at baseline. Thirty-five percent of 313 subjects with an initial positive PPD had a negative PPD test at follow-up, 26% of whom had a positive candida skin test at the same time as the negative PPD test. Baseline MTBCF-stimulated IFN-gamma levels were significantly higher among PPD-positive subjects who remained PPD-positive than in those who were falsely negative. We conclude first that anergy is unstable and second that anergy testing is unreliable in identifying HIV-infected adults who are not infected with MTB and should not be used routinely for this purpose in assessing indications for isoniazid preventive therapy.     

Immunosuppressive properties and circulating life of Achromobacter glutaminase-asparaginase covalently attached to polyethylene glycol in man

The immunosuppressive effects and circulating life of Achromobacter glutaminase-asparaginase (GA) covalently attached to polyethylene glycol (PEG) were examined in human subjects following a single iv dose of 1000 IU/m2. Plasma half-life of PEG-GA was 72 hours. Skin test reactivity to recall antigens (mumps and tuberculin) was lost in all four patients tested. In vitro phytohemagglutinin-induced blastogenesis, "natural killing," and phytohemagglutinin-induced cell cytotoxicity was diminished as long as enzyme levels were detectable. In vivo and in vitro activities returned to normal following total plasma clearance of enzyme.     

Identification of beta-L-gulose as the sugar moiety of the main polar lipid Thermoplasma acidophilum

We studied the antibody binding capacity (ABC) of various cell-surface antigens in normal human fetuses and term neonates on lymphocyte, monocyte, and polymorphonuclear (PMN) cells by quantitative flow cytometry also designated by quantimetry. Analysis of changes of expression level on these leukocytes during the developmental process was also investigated. The results indicated that the ABC values of most studied markers change during the maturational process. The ABC of lymphocyte-associated antigens studied such as CD5 and CD7 showed only a decrease from fetus to adult, whereas according to the type of molecule on monocyte and PMN there was either an increase or a decrease of ABC values dependent on the stage of the developmental process, from fetus to neonate or from neonate to adult. However, the ABC values of leukocyte membrane antigens such as CD16, CD46, and CD55 on all leukocytes and CD11b, CD11c, and CD35 on myeloid cells did not change. Their expression level was already mature in fetuses compared with adult cells. In addition, in this quantimetric approach, the analysis of the results for CD11a and CD8 suggested that the changes of CD11a expression level on lymphocyte subsets can depend on one mechanism, whereas there are probably at least two for CD8. Furthermore, the expression patterns of CD5, CD7, and CD11a change during maturation. We concluded that, even if the neonate response pattern to immunological challenge differs from an adult and this is based primarily on the relative numbers and functional activity of lymphocyte T subsets (especially TH1/TH2) and their cytokine profiles, these quantitative and qualitative phenotypical differences might also contribute to explain the functional peculiarities of leukocyte fetal and cord blood cells. All these findings support the notion of immaturity and maturity of ABC expression.     

Comparison of two methods for the assessment of delayed-type hypersensitivity skin responses in patients with human immunodeficiency virus infection

We compared two techniques for detecting delayed-type hypersensitivity (DTH) skin responses in 359 patients infected with human immunodeficiency virus (HIV) (mean CD4+ lymphocyte count, 387/microL). DTH responses were assessed with use of two antigenic panels administered simultaneously: tuberculin purified protein derivative (PPD) plus three control antigens (Candida albicans, mumps antigen, and tetanus toxoid) administered by the Mantoux method and by a multiple-puncture device delivering seven antigens percutaneously (MULTITEST CMI; Institut Mérieux, Lyon, France). Eighty-three patients (23%) were anergic, 216 (60%) reacted to both panels, 55 (15%) did not react to MULTITEST CMI but did react to the antigens administered by Mantoux method, and only five (1%) reacted to MULTITEST CMI without reacting to antigens administered by the Mantoux method (P < .001, McNemar's test). Each of the three possible combinations of PPD plus two control antigens administered by the Mantoux method were also superior to MULTITEST CMI for classifying patients as nonanergic (P < .001, McNemar's test). We conclude that the application of antigens by the Mantoux method is more efficient than MULTITEST CMI for detecting DTH skin responses in HIV-infected patients.     

Proceedings: An investigation into the immunosuppressive properties of oestrogen

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.