Quantification and origin of the immunoglobulins in porcine respiratory tract secretions.

The immunoglobulin content of porcine nasal, tracheal and bronchio-alveolar secretions was determined. IgA was the predominant immunoglobulin in nasal and tracheal secretions, there being no significant difference (P greater than 0.01) between their IgA:IgG ratios of 2.7:1 and 2.4:1, respectively. In contrast, IgG was the predominant immunoglobulin in bronchioalveolar secretions, the IgA:IgG ratio being 0.7:1. The ability of tracheal and lung tissue to synthesize IgA and IgG was determined in vitro. Tracheal cultures synthesized more IgA than IgG whereas in the lungs the reverse occurred, the ratio of IgA:IgG synthesis being 1.4:1 and 0.4:1, respectively. The proportion of serum-derived IgA and IgG in respiratory tract secretions was determined by measuring the transfer of 125I- and 131I-labelled immunoglobulins from serum. It appeared that more than 97% of the IgA was produced locally. More significantly a proportion of the IgG was also synthesized locally. This proportion was smallest in nasal secretions (21.8%) and greatest in bronchio-alveolar secretions (62.6%). The results suggest that IgG, a large proportion of which is locally synthesized, is important in the homeostatic mechanisms of the lower respiratory tract.     

Detection of infectious human immunodeficiency virus type 1 in female genital secretions by a short-term culture method

Infectious human immunodeficiency virus type 1 (HIV-1) is difficult to detect in female genital secretions by standard virus culture techniques. To improve detection of cell-free HIV-1 in female genital secretions, we adapted a short-term assay that uses the multinuclear-activation galactosidase indicator (MAGI) assay. When vaginal lavages from HIV-1-infected women were tested with the adapted MAGI assay, 25 (64%) of 39 lavages with detectable, cell-free HIV-1 RNA were shown to have infectious virus. No infectious virus was found in 10 vaginal lavages from HIV-1-infected women with undetectable vaginal viral loads. Significantly (P < 0.01) more lavages from HIV-1-infected women tested positive for infectious virus by the MAGI assay than by standard peripheral blood mononuclear cell (PBMC) coculture, which detected infectious virus in only 6 (17%) of 35 vaginal lavages. Lavages with viral loads of >10,000 copies per lavage yielded significantly (P < 0.01) more positive cultures than those with <10,000 copies by using the MAGI assay. Detection of infectious HIV-1 in vaginal lavages was not associated with the presence of genital tract infections or CD4(+)-T-cell counts. However, although the results were not significant (P = 0.08), the MAGI assay detected infectious virus from more vaginal lavages at a vaginal pH of >/=4.5 than at a pH of <4.5. These results indicate that the MAGI assay is more sensitive than PBMC culture methods for detecting infectious virus in female genital secretions. Accurate measurements of infectious virus in genital secretions will improve studies that evaluate sexual transmission of HIV-1.     

Evaluation of a quantitative competitive PCR assay for measuring herpes simplex virus DNA content in genital tract secretions.

Previous studies have shown an association between the approximate titer of herpes simplex virus (HSV) DNA in clinical specimens and the ability to isolate HSV from genital secretions. To control for variance in amplification conditions, we developed a competitive quantitative PCR (QC PCR) for the detection of HSV DNA. The assay accurately measured from 10 to 10(6) copies of HSV DNA. We compared the QC PCR with our previous semiquantitative detection method and found concordance for 61 of 63 positive specimens. We also evaluated the HSV DNA content from individual swabs of genital secretions obtained from individual sites of the genital tract (cervix, vulva, and rectum) with that from one swab with secretions from all three sites. The concordance for detecting HSV DNA was 91%; for only 4 of 143 collection days was there a > 1 log difference between the two collection methods. A single swab with secretions from all three genital sites and evaluated in a QC PCR format can accurately measure the frequency of subclinical and clinical shedding of HSV and the titer of HSV shed from the genital region. Such an approach should be very useful in the evaluation of antiviral chemotherapy for HSV.     

The origins of the immunoglobulins in the mucous secretions of cattle

The predominating immunoglobulin in the mucous secretions of cattle appears to be IgG1. Radioactive tracer studies using ¹³¹I-labelled IgG1 and ¹²⁵I-labelled IgG2 suggest that the larger part of this immunoglobulin is not derived from the plasma but is locally synthesized. Immunofluorescence studies indicate that this immunoglobulin is produced by lymphoid cells which can be observed in the lamina propria and at the base of the villi in the mucosae of the gut and respiratory system. There appears to be some degree of selectivity of transport of IgG1 from the plasma compared with IgG2, although this varies with the tissues.     

Validation of a method to establish practice-based stroke and TIA registers.

This study compares two methods to establish stroke and transient ischaemic attack (TIA) practice-based registers, which are of particular relevance to practices with limited diagnostic coding. Both arms involved a notes review of all patients taking antiplatelets or anticoagulants, and, either a further notes review of all patients with ischaemic heart disease (IHD) or diabetes (extensive arm), or asking about a history of stroke or TIA during IHD or diabetic clinics (pragmatic arm). The extensive arm involved searching 11% of the practice notes, whereas the pragmatic arm only involved 3% and had almost as high a yield. This study suggests that the pragmatic method could be used to help build practice-based stroke and TIA registers.     

The origin of immunoglobulins in semen

IgG and IgA, albumin and lactoferrin as well as the semen compartment parameters acid phosphatase, fructose and spermatozoa were determined in separately collected fractions of the same ejaculate of some normal donors. The distribution over the fractions per ejaculate of IgG, IgA and albumin was generally more or less similar to the distribution of acid phosphatase indicating that the bulk of these proteins enters the semen via the prostate and not via the vesicles or testis and epididymis. The distribution of lactoferrin unexpectedly was not clearly related to fructose. IgM could not be detected.

The concentrations in the (eight) total ejaculates expressed as percentages of the serum concentrations were for albumin slightly higher than for IgG, both in the order of 1% and moreover correlated with each other, indicating that IgG reached the seminal fluid in general by transudation from the circulation. The relative IgA concentrations could not be measured exactly but seemed to be slightly higher than of albumin, and not correlated to albumin concentrations, suggesting that local production of IgA may occur also.

     

Coccidioidomycosis of the female genital tract

Female genital tract involvement is a rare manifestation of disseminated coccidioidomycosis; to our knowledge, only ten patients have previously been described in the English literature. We describe a patient who seems to be unique in that she developed female genital tract coccidioidomycosis and coccidioidal peritonitis after chemotherapy for Hodgkin's disease. Coccidioidomycosis of the female genital tract is usually manifest as granulomatous endometritis and/or granulomatous tubo-ovarian disease with peritonitis. The diagnosis of coccidioidomycosis was unsuspected clinically in all 11 reported cases (including our patient); initial diagnosis was made by biopsy or culture in all 11 patients. In eight of the reported cases of female genital tract coccidioidomycosis (including our patient), clinical improvement occurred after treatment with surgery or antifungal chemotherapy; three patients died of disseminated coccidioidomycosis.     

女性生殖道黄色瘤样肉芽肿性炎

目的 探讨女性生殖道黄色瘤样肉芽肿性炎的病因、临床病理学特征和生物学行为.方法 对3例女性生殖道黄色瘤样肉芽肿性炎作了临床病理观察、免疫组化染色,并做了随访.结果 3例女性生殖道黄色瘤样肉芽肿性炎1例累及卵巢,1例累及双侧输卵管,1例累及子宫内膜.年龄分别是32、22和62岁.组织学观察显示病灶内大量泡沫状的组织细胞,混有淋巴细胞、浆细胞和少量嗜中性粒细胞浸润,偶见多核巨细胞.免疫组化染色:泡沫细胞CD68阳性,淋巴细胞CD3和CD20阳性,κ、λ阳性.结论 女性生殖道黄色瘤样肉芽肿性炎虽是一种罕见的炎性病变,但只要记住它的存在及特点,就可以避免误诊.     

Endocrine cells in the female genital tract

Endocrine cells are normal inhabitants of the para-urethral, Bartholin's and endocervical glands and of mesonephric rests. All these cells were characterized as serotonin-storing cells. In the para-urethral and Bartholin's glands, serotonin-containing cells were most often found in the transitional epithelium of excretory ducts. Endocrine cells participated in some pathological conditions. Abundant argentaffin cells were observed among the terminal ductules in chronic bartholinitis and serotonin-storing cells were identified in a peculiar ectocervical epithelium. Numerous serotonin-storing cells were detected in a well-differentiated adenocarcinoma of cervix occurring in a patient with the Peutz-Jeghers syndrome. Argyrophilic cells were present in cases of endometrial carcinomas; a striking feature was the demonstration of gut peptide hormones in an unusual type of endometrial adenocarcinoma. Finally, serotonin-storing cells were a constituent of Brenner tumours. It is suggested that a similar endocrine pattern may be shared by tissues originating from both Müllerian ducts and the urogenital sinus.     

Screening of high cytotoxic tumor killer cells using a sensitive adherent target detachment assay

Screening of high cytotoxic tumor killer cells is of great importance in adoptive immunotherapy. Here, we describe a more sensitive assay, as compared to traditional 51Cr- or Calcein-release assay, to measure cytolytic activity of killer cells. This adherent target detachment (ATD) assay is carried out in microwells of Terasaki tissue culture trays using adherent tumor cells as targets. Target tumor cells are seeded at the concentration of 300-400 cells/well and incubated overnight to allow for the adhesion of cells to the plastic surface of the wells. Effector cells were added at various effector: target (E:T) ratios, and incubated for 24 h. During incubation, dead target cells became nonadherent and together with the added effector cells were removed by washing. The remaining viable adherent target cells were stained with acrydine orange contained in the quencher and optically counted by microcomputer. A notable dose-dependent killing on target tumor cells was reproducibly obtained by tested effector cells. Cytotoxic activities of most effector cells were significantly higher in the 24-h ATD assay than those of concordant 4-h target cell lysis test. Chloroquine (chq) inhibition test in the 24-h ATD assay showed negative or weak inhibition on cytotoxicity against adherent targets. Linear regression analysis also manifested the lack of a close correlation between 4- and 24-h target cell lysis tests, indicating these two assays measured different killing activities of the same effector cells. Because of its high sensitivity and reproducibility, this semiautomatic 24-h assay with computerized fluorescence measurement system could serve as a sensitive screening assay to select high cytotoxic tumor killer cells in adoptive immunotherapy.     

Contribution of immunoenzymatic technics to the assay of anti-native DNA antibodies in the biological diagnosis of lupus erythematosus disseminatus

Anti native DNA antibodies (anti nDNA Ab), which are a highly specific feature of systemic lupus erythematosus (SLE) were measured by 3 methods: an enzyme linked immunosorbent assay (ELISA), an indirect immunofluorescence test on Crithidia luciliae (IFCL) and the Farr assay (reference test). 114 sera from patients with SLE or another connective tissue disease or without autoimmune rheumatic disease were tested. This study showed that ELISA seemed to be a more sensitive and specific test than IFCL (classical test). ELISA was also as sensitive as the Farr assay. ELISA should replace IFCL for the diagnosis and the follow up of patients with SLE. In other connective tissue diseases, ELISA might give more positive results. Thus these had to be confirmed, especially in the case of low antibodies levels, by using another method (e.g., the Farr assay).     

A simple and rapid immunoenzymatic assay of prostatic acid phosphatase in serum

A method is described for the quantitative assay of human prostatic acid phosphatase after its isolation from serum by a double antibody technique employing a specific rabbit antiserum and a goat anti-rabbit gammaglobulin serum. After centrifugation, the precipitate is analyzed for phosphatase activity using p-nitrophenylphosphate as substrate. The method allows a rapid, precise, specific and sensitive assay.     

Rapid detection of cytomegalovirus DNA in urine samples with a dot blot hybridization immunoenzymatic assay.

A dot blot hybridization immunoenzymatic assay for the rapid detection of cytomegalovirus DNA in urine samples was developed by using a digoxigenin-labeled probe which was immunoenzymatically visualized by antidigoxigenin Fab fragments labeled with alkaline phosphatase. A total of 516 urine samples from different groups of subjects were analyzed, and the hybridization assay was able to yield results within 24 h. The results obtained were compared with results for detection of cytomegalovirus antigens in infected cell cultures.