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Effect of normal and immune sera on Haemophilus ducreyi 35000HP and its isogenic MOMP and LOS mutants.

A bactericidal assay was developed in order to test the effect of hyperimmune rabbit sera on the viability of serum-resistant Haemophilus ducreyi 35000HP. Testing of several lots of rabbit complement and time course experiments showed that the serum-sensitive H. ducreyi CIPA77 was killed efficiently by 25% complement at 35 degrees C in 3 h. We hypothesized that incubation of 35000HP under these conditions with the appropriate bactericidal antibody would kill this strain. A panel of high titre rabbit antisera was developed and tested against 35000HP. The panel included antisera raised to whole cells, total membranes, Sarkosyl-insoluble outer membrane proteins, the H. ducreyi lipoprotein, and the peptidoglycan-associated lipoprotein. None of the antisera convincingly showed bactericidal activity. The bactericidal assay was also used to determine the effect of normal human serum (NHS) on isogenic mutants of 35000HP. 35000HP-RSM2, an Omegakan insertion mutant that expresses a truncated lipooligosaccharide, was as resistant to NHS as its parent. A mutant deficient in expression of the major outer membrane protein (35000. 60) was sensitive to NHS. We conclude that 35000HP is relatively resistant to normal and hyperimmune sera, and that the major outer membrane protein contributes to this resistance.

Animals↗

Toxicity and immunogenicity of purified Haemophilus ducreyi cytolethal distending toxin in a rabbit model.

The cytolethal distending toxin of Haemophilus ducreyi (HdCDT) is a three-component toxin that induces the arrest of the mammalian cell cycle in the G2 phase. All of the individual gene products, CdtA, CdtB and CdtC, are required for toxic activity on cultured mammalian cells. The CdtB component alone exerts nuclease activity. The individual HdCDT components were purified by affinity chromatography or ion-exchange chromatography followed by gel-filtration. HdCDT was reconstituted and purified by the immobilization of a GST-CdtB fusion on a GSTrap column and the subsequent addition of cell sonicates from Escherichia coli recombinants that produced CdtA and CdtC. The purified HdCDT preparation contained all three CDT proteins, as detected by immuno-blotting, and had high cytotoxic activity (10(6)CPU/ml). Immunization of rabbits with the HdCDT complex and with the individual CdtA, CdtB and CdtC proteins elicited high titres of antibodies, as detected by ELISA. All of the immune sera had toxin-neutralizing activities. The pathological effects of the HdCDT complex were investigated in rabbits, since the proliferation of two rabbit cell lines, SIRC and RK-13, was inhibited by HdCDT. Intradermal injection of HdCDT (1, 10, 50 and 100microg protein) into naive rabbits resulted in dose-dependent skin reactions (erythema) about 24h after injection. Similar effects were not observed when the individual HdCDT proteins were injected. HdCDT injection into immune rabbits resulted in dose-dependent skin responses that were characterized by both erythema and oedema. Histological evaluation of the 24-h lesions in naive rabbits that were injected with HdCDT, revealed moderate levels of inflammatory cells, which were mainly granulocytes and macrophages, and dilatation of blood vessels. The skin reactions in HdCDT-injected immunized rabbits showed pronounced vascular changes and extensive infiltration of inflammatory cells, including eosinophils. All of the pathological changes healed after 3 days. In conclusion, purified HdCDT holotoxin is a complex of all three CDT proteins and all three components induce neutralizing antibodies when injected in rabbits. HdCDT causes dose-dependent pathologic skin reactions in both naive and immune rabbits, which is characterized by increased inflammatory responsiveness after each immunization.

Animals↗

AIDS and heterosexual anal intercourse.

Heterosexual anal intercourse is rarely discussed in the scientific literature. Review of the literature suggests the silence is linked to ethnocentric discomfort about it among researchers and health care providers, coupled with the misconception that anal sex is a homosexual male practice, not heterosexual. Review of surveys of sexual practices suggest that heterosexual anal intercourse is far more common than generally realized, more than 10% of American women and their male consorts engaging in the act with some regularity. Sexually transmitted disease (STD) data, especially where only the rectum is infected with gonorrhea or other STD agents, buttresses survey data. Considerably more heterosexuals engage in the act than do homosexual and bisexual men, not all of whom participate in anal coitus. Anal intercourse carries an AIDS risk for women greater than that for vaginal coitus, just as receptive anal intercourse carries a very high risk for males. Infection with the AIDS virus is increasingly documented in women engaging in anal coitus with infected males, in America, Europe, and Latin America. Women in Western countries are less likely to continue HIV infectivity chains than are males engaging in same-gender anal intercourse.

Acquired Immunodeficiency Syndrome↗

[Recurrent chancriform mucous membrane ulcer in plasmacytoma with secondary IgA deficiency. Pyoderma chancriforme of the tongue].

A 66 year old patient presented with a nine month history of recurrent oral ulcerations involving the tongue. We diagnosed chancriform pyoderma and a previously not identified multiple myeloma with secondary immunoglobulin deficiency. Clinically and histologically we excluded a necrotizing ulcerative stomatitis as found in individuals with cellular immunodeficiency as in late-stage HIV-infection. On culture only Neisseria catarrhalis was found. Chancriform pyoderma is often associated with local bacterial infections, especially Staphylococcus aureus. The most common sites are the genitalia and periorbital region; involvement of the oral mucosa is uncommon. To the best of our knowledge, this is the fourth reported case with tongue lesions. The multiple myeloma-associated immunoglobulin deficiency might have facilitated the oral manifestations of chancriform pyoderma.

Aged↗

The office approach to the sexually transmitted diseases: Part II.

All physicians share a responsibility for educating the public in the expanding risks of sexually transmitted diseases, and in the vital importance of safe sexual practices. Both the public health implications of undiagnosed STDs and the adverse social consequences of an incorrect diagnosis emphasize the need to comprehensively and precisely diagnose these diseases, despite the problems encountered in obtaining material for culture. Current interest in AIDS should not obscure the facts that genital herpes continues to increase in prevalence, and that syphilis no longer is declining. The many extragenital syndromes associated with STDs, such as perihepatitis, polyarthritis, and acute septic states make it necessary to evaluate all sexual contacts.

Acquired Immunodeficiency Syndrome↗

Detection of Haemophilus ducreyi lipooligosaccharide by means of an immunolimulus assay.

A murine monoclonal antibody (MAb) directed against a surface-exposed epitope of the lipooligosaccharide (LOS) of Haemophilus ducreyi strain 35000 was shown to be reactive with all 37 strains of this pathogen tested in a colony blot-radioimmunoassay. The LOS epitope bound by this MAb appeared to be stably expressed by H. ducreyi growing in vitro. The use of this MAb in the immunolimulus system revealed that it could detect purified H. ducreyi LOS at a level of 25 pg/ml. Similarly, this immunolimulus system could detect as few as 1000 colony forming units of in vitro-grown H. ducreyi cells per ml of buffer. When this MAb was utilized in the immunolimulus system together with lesion material from rabbits infected with two different H. ducreyi strains, a positive reaction was obtained with every sample tested, even when no viable organisms were present in the lesion material. In contrast, this MAb yielded consistently negative results when used in the immunolimulus system with lesion material from animals infected with Staphylococcus aureus.

Animals↗

Oculogenital disease.

This review comprises brief discussions of the epidemiology, clinical manifestations, laboratory features, and treatment of the oculogenital diseases as they appear typically in the newborn infant, the child, and the adult. Inclusion conjunctivitis is the most frequently occurring oculogenital infection in all three categories of patient. Oculogenital diseases are becoming increasingly important in both frequency and severity. They can be extremely serious and may thus require the immediate attention of the physician.

Adult↗