Cross-reactive cytotoxic T lymphocytes against human immunodeficiency virus type 1 protease and gamma interferon-inducible protein 30.

The gamma interferon (IFN-gamma)-inducible protein 30 (IP-30) signal peptide -11 to -3 (LLDVPTAAV) is a prominent self peptide expressed with the class I human histocompatibility leukocyte antigen A2 (HLA-A2). Stimulation of peripheral blood mononuclear cells (PBMC) from HLA-A2 human immunodeficiency virus type 1 (HIV-1)-infected individuals with an HLA-A2-restricted HIV protease (PR) peptide 76-84 (LVGPTPVNI) activated cytotoxic T lymphocytes (CTL) against the IP-30 signal peptide. Since HIV-1 PR 76-84 stimulated CD8+ T cells from these individuals to secrete IFN-gamma, we tested whether the activation of IP-30-specific CTL in vitro resulted from T-cell cross-reactivity or from up-regulation of IP-30 by IFN-gamma. Neither high levels of exogenous IFN-gamma nor incubation of PBMC with other HIV peptides triggering substantial IFN-gamma release activated IP-30-specific CTL. Although the IP-30 signal peptide did not stimulate IFN-gamma release from freshly isolated PBMC, it activated CTL in vitro against itself and HIV PR 76-84. Peptide-stimulated IFN-gamma release, cold target inhibition, and HLA-A2/immunoglobulin dimer-mediated binding and depletion of effector cells all indicated that in vitro stimulation with HIV PR 76-84 or the IP-30 signal peptide activated a comparable population of cross-reactive effector cells. Neither IP-30 nor HIV PR 76-84 activated CTL against themselves following in vitro stimulation of PBMC from non-HIV-infected HLA-A2 individuals. Peptide titrations indicated higher-avidity T-cell interactions with HIV PR 76-84 than with the IP-30 signal peptide. These data indicate that HIV PR 76-84 is a heteroclitic variant of the IP-30 signal peptide -11 to -3, which has implications for immune memory and autoimmunity.

Fas/FasL-independent activation-induced cell death of T lymphocytes from HIV-infected individuals occurs without DNA fragmentation.

We assessed the effects of activation with phorbol myrystic acetate (PMA) and ionomycin on peripheral blood mononuclear cells (PBMC) from HIV-infected individuals by (51)Cr release, propidium iodide (PI) uptake, electron microscopy, and DNA analysis. Up to 70% (51)Cr release was induced from PBMC of HIV-infected individuals, versus up to 26% (51)Cr release from PBMC of non-HIV-infected volunteers. Flow cytometry identified mostly T cells undergoing activation-induced cell death (AICD). The kinetics of (51)Cr release and the effects of cold target inhibitors were consistent with cell-mediated cytotoxicity. Certain anti-CD3 antibodies or extracellular Ca(2+) chelation prevented AICD, but antagonistic anti-Fas antibodies, caspase inhibitors, and cycloheximide had no effect. The antioxidants thiourea and N-acetylcysteine reduced AICD, indicating a role for oxidative stress. Electron microscopy revealed plasma membrane disruption with nuclear integrity, while DNA analysis showed intact chromosomal DNA. This form of T cell AICD triggered by PMA and ionomycin differs from classical apoptosis in the absence of either caspase involvement or DNA fragmentation.

HIV-1 DNA burden in peripheral blood CD4+ cells influences disease progression, antiretroviral efficacy, and CD4+ T-cell restoration.

Integration of human immunodeficiency virus type-1 (HIV-1) proviral DNA into host cell genomic DNA ensures viral persistence despite suppression of active replication. Because HIV RNA originates from integrated HIV DNA, HIV RNA and DNA loads should interrelate when suppression of viral replication is incomplete. In addition, the link between proviral DNA formation and generation of HIV-1 genetic diversity suggests that the ease with which HIV escapes immune or drug-based suppression should vary with proviral load. Thus, HIV proviral load should have unique prognostic significance independent of the highly labile plasma HIV RNA levels commonly used to monitor patient status. To test this possibility, we developed a simple standardized research assay estimating the proportion of CD4+ peripheral blood mononuclear cells (PBMC) carrying HIV-1 DNA and investigated associations between this parameter, plasma virus load, long-term efficacy of antiretroviral therapy and restoration of CD4+ T cells. Lower proportions of CD4+ PBMC carrying HIV-1 DNA were associated with lower peak plasma HIV RNA levels and with more favorable long-term responses to antiretroviral therapy. These results suggest that HIV proviral load affects both disease progression and responsiveness to antiretroviral therapy. Therefore, new anti-HIV therapies addressing the stable pool of HIV proviral DNA should be developed to improve long-term prospects for suppression of HIV replication.

Functional and genetic integrity of the CD8 T-cell repertoire in advanced HIV infection.

BACKGROUND: HIV-specific cytotoxic T lymphocytes (CTL) can restrict HIV replication in acute and chronic infection, but disease progression occurs in parallel with declining CTL activity. An understanding of why CTL fail to control HIV replication might reveal important mechanisms of disease progression and enhance prospects for developing effective CTL-based immunotherapies. OBJECTIVES: To investigate the functional integrity, T-cell repertoire diversity, and HIV reactivity of CD8 T lymphocytes in individuals with advanced HIV infection. METHODS: Individuals were considered to have progressed to advanced HIV infection if their total T-cell count was < 500 x 10(6) cells/(l) on at least two successive clinic visits. CD8 T cells from these individuals were analyzed for CTL function, HIV reactivity and T-cell receptor (TCR) diversity by chromium release assays and reverse transcriptase polymerase chain reaction. RESULTS: CD8 T cells from all individuals with advanced HIV infection proliferated and differentiated into functional CTL in vitro. Despite extremely low T-cell counts and previous AIDS-defining illnesses, six individuals had inducible anti-HIV CTL responses. In two additional cases, HIV-specific CTL activity became detectable following significant treatment-associated remission of T-cell lymphopenia. Assessment of TCRbetaV gene family representation and betaV gene intrafamily diversity indicated CD8 T-cell repertoire diversity is maintained through advanced HIV infection. CONCLUSIONS: These data suggest that HIV-specific CTL activity can be selectively compromised while the functional and genetic integrity of the CD8 population as a whole remains intact. A substantial fraction of individuals retain inducible anti-HIV CTL activity through advanced HIV infection and, in at least some cases, effective treatment can restore HIV-specific CTL responses even at this late stage of disease.

Accreditation of postgraduate medical education in the United States and Canada: a comparison of two systems.

The systems for accrediting residency programmes in the United States and Canada, although they have developed independently in the two countries, have similar objectives and accreditation requirements. Both have become increasingly focused over the past several decades on the importance of educational programmes structured to provide graded professional responsibility with appropriate guidance and supervision to residents according to their level of training, ability and experience. The Canadian model used by the Royal College of Physicians and Surgeons of Canada is a centrist one, with accreditation decisions on all programmes in all specialties being vested in a single, multidisciplinary accreditation committee. The American model developed by the Accreditation Council for Graduate Medical Education is a distributive one, with accreditation decisions being vested in each specialty Residency Review Committee. In both models, accreditation is based on a system of periodic on-site visits during which both faculty and residents are interviewed by a surveyor to provide the accrediting body with a first-hand evaluation of the extent to which each accredited programme meets the standards of accreditation. While they are similar in purpose, there are significant differences in the operation of the two systems.

Single-dose ceftriaxone for chancroid.

Men with genital ulcers that were culture positive for Haemophilus ducreyi were treated with intramuscular ceftriaxone and randomized to three different dose regimens. All but 1 of 50 men treated with 1 g of intramuscular ceftriaxone were cured. Similarly, 0.5 and 0.25 g cured 43 of 44 men and 37 of 38 men, respectively. A single dose of 250 mg of intramuscular ceftriaxone is an effective treatment for chancroid.

Factors affecting Staphylococcus epidermidis growth in peritoneal dialysis solutions.

Staphylococcus epidermidis is the most frequent cause of peritonitis complicating continuous ambulatory peritoneal dialysis. We studied factors that might influence the growth of S. epidermidis in commercially available peritoneal dialysis solution (PDS). Test strains were inoculated into PDS and incubated overnight at 37 degrees C. Samples were removed at appropriate intervals, bacterial counts were performed, and growth curves were constructed. We studied the effects of various osmolarities, the neutralization and acidification of fresh and spent PDS, and the effect of intraperitoneal dwell time on the ability PDS to support growth of S. epidermidis. In fresh PDS, numbers of bacteria remained constant after 24 h. No significant differences in growth were observed among PDS with 0.5, 1.5, 2.5, and 4.25% glucose. Neutralizing acidic fresh PDS had no effect on bacterial growth. However, growth did occur in spent PDS. PDS which was recovered after only 2 h in the peritoneal cavity supported growth to the same extent as did PDS recovered after 4 to 6 h. Mean log10 changes after 24 h of incubation were as follows: for fresh PDS, -1.3; after 2 h dwell time, 2.9; after 4 h dwell time, 1.9; and after 6 h dwell time, 1.3. Acidification of spent PDS to less than pH 6.35 produced less rapid growth; mean log10 increases after 24 h of incubation were 1.9 for pH 7.75, 1.6 for pH 6.35, 0.6 for pH 5.75, and 0.7 for pH 4.95. Fresh PDS of all available osmolarities neither supported the growth of S. epidermidis nor was bactericidal. Spent PDS supported bacterial growth, and this growth was partly independent of the neutralization which occurred during the dialysis.

Von Hippel-Lindau disease in a Newfoundland kindred.

Von Hippel-Lindau disease, an autosomal dominant condition with complete penetrance, has been recognized in a large family that originated in Newfoundland but has some members who live in New Brunswick and Ontario. A collaborative investigation was begun in 1982 to document the number of affected members and the extent of their disease and to improve management of the disease. The condition has been documented in 38 members of the family, 28 living and 10 dead. The most common manifestations are retinal angioma (present in 60% of the gene carriers) and pheochromocytoma (present in 53%). Of the 28 living affected members 14 had been identified before the study began. Only 3 of the 14 patients in whom the disease was subsequently diagnosed presented with symptoms; in the remaining 11 the condition was detected by routine screening. Overall the mean age at the time of diagnosis was 23 years; in the 21 affected members of the fourth generation it was 18 years. The authors outline a regimen of regular screening for members at risk that has evolved as a result of their experience with this family.

Herpes simplex virus in clinically suspected chancroid in Nairobi, Kenya.

PIP: Of 110 males selected for review with possible chancroid, 96 were clinically diagnosed as having chancroid, 7 as having herpetic lesions, and 7 as having syphilis. Of the 96 patients diagnosed clinically as chancroid, 76 (79.2%) were culture positive for H. ducreyi. 9 (9.4%) of these 96 patients yielded Herpes Simplex Virus (HSV). Both HSV and H. ducreyi were isolated from 5 of the patients, and from 4 of the patients HSV alone was isolated. 7 patients (6.4%) were clinically diagnosed as having herpetic ulcers. 5 of these grew HSV. Overall, 14 of the 110 patients (12.7%) yielded HSV. 1 patient, who presented with small vesicular lesions characteristic of HSV, yielded the virus on culture. The vesicles were initially negative for H. ducreyi, but 6 days later he had developed deep purulent ulcers in the same sites as the vesicular lesions and became culture positive for H. ducreyi snd HSV-negative. The possible association between HSV and chancroid is discussed in the light of these findings and comparisons made between the results of the present study and earlier findings made in Kenya and elsewhere, with suggestions being given as to the reasons for the apparent differences. The HSV isolation techniques used in this study may be less sensitive than those used in other studies, but it is highly unlikely that this possibility alone accounts for all of the observed differences. Patients with hepetic ulcers may be less likely to present early in the course of the disease, if at all, believing the infection to be minor and one that will heal on its own. It is also possible that HSV infection is less common in Kenya, either alone or as an initiator of chancroid, than in the US or Europe, becuase of a higher rate of childhood HSV infections in Kenya, which may confer a degree of immunity against genital HSV infection in this population. The lower prevalence of HSV in association with H. ducreyi reported may be at least partly the result of a much higher incidence in Kenya of chancroid which is not initiated by HSV. A higher incidence of HSV genital infection in Europe and America would also make it more likely that HSV would fortuitously be isolated more frequently from H. ducreyi positive lesions.^ieng

Treatment of pneumonia and other serious bacterial infections with cephapirin.

Nineteen patients with pneumonia due to Streptococcus pneumoniae, Staphylococcus aureus, Klebsiella pneumoniae, or Escherichia coli were treated with 4 to 18 g of cephapirin daily. There were three treatment failures. One patient each with pneumonia due to E. coli or S. pneumoniae died despite apparent eradication of the pathogen. Lobar pneumonia due to K. pneumoniae progressed during therapy in a third patient to lung gangrene, necessitating pneumonectomy. Five additional patients with pneumococcal pericarditis or septic bursitis, empyema, cannula-associated bacteremia, and thoractomy wound infection due to S. aureus were cured. All isolates of S. aureus, S. pneumoniae, and group A Streptococcus were inhibited by 0.8 mug of cephapirin per ml; minimal inhibitory concentrations of cephalothin were similar. Ninety percent of K. pneumoniae, 85% of Proteus mirabilis, 73% of E. coli, and 30% of Enterobacter were inhibited by 12.5 mug cephapirin per ml. All isolates of Pseudomonas, Serratia and indole-positive Proteus had a cephapirin minimal inhibitory concentration of [Formula: see text] 100 mug/mg. Serum concentrations after intravenous and intramuscular injection were similar to those reported for cephalothin. The intramuscular injections were moderately painful, and intravenous infusions caused phlebitis in three of nine patients treated with doses up to 18 g per day. Cephapirin appears comparable to cephalothin in vitro and is an effective agent in treatment of infection due to S. aureus and S. pneumoniae.