HIV-1 and HIV-2 infection associated with AIDS in Abidjan, Côte d'Ivoire.

In September 1987, a seroprevalence study of HIV-1 and HIV-2 infection was conducted among 956 people from different groups in Abidjan, Côte d'Ivoire. Groups examined were hospitalized patients (Internal Medicine and Infectious Disease Departments, Centre Hospitalier Universitaire de Treichville, Abidjan), outpatients at tuberculosis treatment centers, blood donors, women attending an antenatal clinic, and patients attending sexually transmitted disease (STD) clinics. Total HIV infection prevalence ranged from 10% in STD clinic patients and pregnant women to 45% in hospitalized patients on an infectious diseases service. Within groups, HIV-1 infection was 2-6.5 times more prevalent than infection with HIV-2. One-quarter of HIV-seropositive people were serologically reactive to both HIV-1 and HIV-2 on enzyme-linked immunosorbent assay and Western blot. Clinical conditions previously observed in patients with HIV-1 infection were observed in people infected with HIV-2 only, as well as in those with HIV-1 infection and dual serologic reactivity. An isolate of HIV-2 was obtained on culture from a person with wasting disease and chronic fever. The results of this study suggest that infection with HIV-1 and HIV-2 is epidemic in Côte d'Ivoire, and that HIV-2 may be associated with AIDS.

Identification and characterization of a cell surface proteoglycan on keratinocytes.

Proteoglycans fill the intercellular space between keratinocytes but their structure and function are not well understood. We have identified and partially characterized one intercellular proteoglycan on human keratinocytes, for which we propose the name epican (epidermal intercellular proteoglycan). Monoclonal antibodies (MoAb) were generated from a mixture of keratinocyte proteoglycans. One, designated MoAb17, identified the core protein of an intercellular proteoglycan that had an apparent mobility of greater than 250 kDa on Western blots. The core protein itself had an apparent mobility of 180 kDa following deglycosylation with trifluoromethanesulfonic acid. Enzymatic deglycosylation revealed that most core protein molecules were substituted with heparan sulfate but that some carried chondroitin sulfate instead. Smaller forms of the core protein were more abundant in tissue-culture medium than in cell extracts. This proteoglycan was localized by immunofluorescence to the intercellular space of the epidermis and the surface of keratinocytes in vitro, particularly at cell-cell contacts. MoAb17 did not react with protoglycans extracted from other skin cells, nor did it bind to basement membranes or connective tissue. Comparison of Western immunoblots using MoAb17 and antibodies to core proteins of other proteoglycans suggested that epican is not related to syndecan but is a member of the CD44 family.

A comparative ultrastructural localization of concanavalin A, wheat germ and Ricinus communis on glomeruli of normal rat kidney.

Three lectins, concanavalin A, peroxidase labeled wheat germ and peroxidase labeled Ricinus communis have been utilized to determine the localization of various saccharide determinants in the glomerulus of the normal rat kidney. In order to obtain a homogeneous penetration of the lectins throughout the whole section, various technical parameters were studied. Only with concanavalin A, a diffuse labeling of the endoplasmic reticulum was obtained. With the three lectins, the basement membrane, mesangial matrix and the cell coat of the three cell types in the glomerulus were labeled. Differences in the labeling of the capillary wall were also noted.

Podocytes in aminonucleoside glomerulonephritis investigated ultrastructurally with concanavalin A.

Peroxidase labeled concanavalin A (Con A) permits the detection of some saccharide determinants. This histochemical technique permits the visualization of cellular pathological modifications not observed with other methods. Its use in an ultrastructural study of nephritis induced by a single injection of aminonucleoside demonstrated the following in podocytes. The Con A positive endoplasmic reticulum (ER), essentially the rough ER, lost its normal linear and network appearance to take on a dot dash pattern. ER contents but not attached ribosomes and membranes were Con A positive. The dot dash pattern, due to a fragmentation of the ER, appeared prior to the onset of proteinuria and was attenuated before the disappearance of proteinuria. These changes of the ER were not observed in other proteinuric states. This suggests that aminonucleoside can damage the synthesis apparatus of podocytes, revealed by the Con A method.

Histochemical and ultrastructural characterization of subendothelial glycoprotein microfibrils interacting with platelets.

The interaction of human blood platelets with collagenase-treated rabbit subendothelium was studied by histochemical ultrastructural methods and by morphometric semi-quantitative analysis. Aortas were deendothelialized and incubated: 1) with a highly purified bacterial collagenase whose specificity was controlled; and 2) with the same collagenase followed by chymotrypsin. For histochemical studies, tannic acid, ruthenium red, and peroxidase-labeled Ricinus communis and concanavalin A were used. Electron microscopy showed that after digestion of fibrillar collagen by collagenase, adherent and aggregated platelets were observed on Ricinus communis-, concanavalin A-, and ruthenium red-positive glycoprotein microfibrils. After successive incubation with collagenase and chymotrypsin, the microfibrils disappeared. No platelets were observed on the remnant amorphous elastin. Morphometric analysis confirmed the interaction of platelets with collagenase-treated subendothelium. In addition, glycoproteins were extracted from collagenase-treated rabbit aortas using 5 M guanidine. Using an in vitro quantitative test, significant platelet adhesion to these glycoproteins was observed. Our results show an interaction between platelets and noncollagenic glycoprotein microfibrils.

Use of a polylysine-saporin conjugate to prevent posterior capsule opacification.

PURPOSE: To determine the feasibility of applying a polylysine-saporin (PLS) conjugate to the lens capsule at surgery to prevent lens epithelial cell (LEC) proliferation and posterior capsule opacification (PCO). SETTING: Department of Research & Development, Bausch & Lomb Surgical, and Department of Ophthalmology, Saint Louis University, St. Louis, Missouri, USA. METHODS: Fluorescein-labeled polylysine was applied to the lens capsule of rabbits after phacoemulsification and analyzed histologically to determine the extent of binding to the lens capsule and surrounding tissues. The cytotoxin saporin was conjugated to polylysine using bifunctional cross-linkers. This PLS conjugate was applied to LECs in culture and to the lens capsules of rabbits. These eyes were monitored for PCO. RESULTS: Polylysine primarily bound to the lens capsule membranes, with little or no binding to surrounding tissues. When PLS was added to LECs in culture, it was internalized and destroyed the cells. Of 9 rabbit eyes treated with PLS during surgery, 1 remained free of PCO for the life of the animal (40 weeks), while 6 showed a delay of cortical regrowth approximately 2 to 3 times that of control eyes. CONCLUSIONS: Polylysine bound selectively to the lens capsule membrane. The PLS conjugation resulted in a toxic agent that targeted the lens capsule and destroyed proliferating LECs. The application of a PLS conjugate during surgery may prevent PCO.

Risk of tuberculosis in patients with HIV-I and HIV-II infections in Abidjan, Ivory Coast.

OBJECTIVE: To examine the association between HIV-II infection and tuberculosis. DESIGN: Cross sectional study comparing the prevalence of HIV-I and HIV-II infections in patients with tuberculosis and in blood donors. SETTING: Abidjan, Ivory Coast, west Africa. PATIENTS: 2043 consecutive ambulant patients with tuberculosis (confirmed pulmonary, presumed pulmonary, or extrapulmonary) and 2127 volunteer blood donors. MAIN OUTCOME MEASURE: Prevalence of HIV-I and HIV-II infections as assessed by presence of serum antibodies. RESULTS: Overall rates of HIV infection were 40.2% in patients with tuberculosis (26.4% positive for HIV-I, 4.7% for HIV-II, and 9.0% for both); and 10.4% in blood donors (7.2% positive for HIV-I, 1.9% for HIV-II, and 1.3% for both). HIV-II infection was significantly more common in patients with all types of tuberculosis than in blood donors (97/2043, 4.7% v 40/2127, 1.9%; odds ratio 3.8%, 95% confidence interval 2.6 to 5.6). CONCLUSION: Both HIV-I and HIV-II infections are associated with tuberculosis in Abidjan. 35% of adult tuberculosis in Abidjan is attributable to HIV infection and 4% specifically to HIV-II.

Butyric acid causes morphological changes in cultured chondrocytes through alterations in the extracellular matrix.

Butyric acid induces characteristic changes in the morphology of chick embryo chondrocytes. Chick embryo chondrocytes when cultured in the absence of butyrate exhibit a spherical morphology and synthesize cartilage-specific chondroitin sulfate proteoglycan (CSPG). When these cultures are initiated and maintained in the presence of butyric acid, chondrocytes exhibit a mesenchymal morphology, a 90% reduction in the synthesis of CSPG, and a 75% reduction in DNA synthesis. The reduced synthesis of CSPG and DNA was shown not to be dependent on the morphological change. Chondrocytes require CSPG in order to express a spherical morphology, since including chondroitinase ABC in the culture media caused the cells to spread. In addition, the treatment of chondrocytes with purified CSPG prior to culture in media containing butyric acid resulted in spherical cells. The butyrate-induced spreading was shown to require either serum or fibronectin and could be prevented with antiserum against chick cell-surface fibronectin (cFn). Cell-surface fibronectin, which was present on both spherical and flattened chondrocytes, organized into fibrils beneath cells which spread. Increased fibronectin synthesis was not responsible for the butyrate-induced morphological change. From this evidence, it is concluded that the mechanism by which butyrate alters the morphology of these cells in culture involves inhibiting CSPG synthesis, thus preventing CSPG accumulation in the extracellular matrix (ECM). The absence of CSPG in the ECM allows fibronectin to mediate spreading of chondrocytes in culture.

Characterization by immunoperoxidase of the lymphocytes with bundle-shaped tubular (BST) inclusions in human normal peripheral blood.

In six normal subjects, no surface immunoglobulins were detected on blood lymphocytes containing bundle-shaped tubular (BST) inclusions, after incubation with anti-human IgM peroxidase-labeled Fab'2 fragments. These Fab'2 fragments from pepsin-digested sheep anti-human immunoglobulins revealed human mu kappa, and lambda chains. These results suggest that the BST inclusions do not belong to B lymphocytes but belong to T cells and/or a third lymphocyte population, including K cells and precursors of T and B cells.

Response of stratified cultures of human keratinocytes to disruption of proteoglycan synthesis by p-nitrophenyl-beta-D-xylopyranoside.

Proteoglycans play a role in regulating proliferation and adhesion of cells to each other and to the basal lamina. Synthesis of proteoglycans is disrupted by beta-xylosides, which serve as alternate substrate sites for glycosaminoglycan chain attachment and therefore prevent glycosylation of the core protein. We have investigated the effects of p-nitrophenyl-beta-D-xylopyranoside (PNP-xyloside) on cultured human keratinocytes. Stratified cultures were incubated for 7 days with PNP-xyloside (0.05-2.0 mM). Concentrations as low as 0.05 mM increased the secretion of free chondroitin sulfate by 10-15-fold over untreated cultures. Cell-associated proteoglycan decreased as PNP-xyloside concentration increased. At 2 mM PNP-xyloside, heparan sulfate as well as chondroitin sulfate addition to core proteins was disrupted: the core protein of epican, a heparan sulfate form of CD44 found on keratinocytes, was detected immunologically but lacked heparan sulfate. 2.0 mM PNP-xyloside reduced the number of attached cells by 20-25% after 7 days, but had little effect on morphology or protein synthesis. These results indicate that intact proteoglycans are not critical for maintaining epidermal keratinocyte stratification, cell-cell adhesion, or growth.

Distribution of blood group antigen A in normal and pathologic human kidneys.

We tested this distribution with an indirect immunofluorescent technique using purified rabbit anti-A antiserum on 21 whole normal kidneys (a group, N equal to 18; AB group, N equal to 1; O group, N equal to 2) and on 349 kidney biopsy samples (A group, N equal to 140; AB group, N equal to 14; O or B group, N equal to 195) representing a large spectrum of renal diseases. In normal kidneys from A and AB groups, the A antigen was detected in the whole vascular endothelium and in the convoluted distal tubules. In secretors, collecting tubules were brightly positive. Epithelial staining was more diffuse in the inner part than it was in the outer part of the medulla. The basement membrane of the inner collecting tubules was positive in frozen sections but not in paraffin sections. In pathologic kidneys, modifications were obvious: (1) The thickened basement membrane of atrophic convoluted distal tubules was brightly stained. (2) Endothelial staining allowed a precise appreciation of the glomerular and interstitial vasculature. (3) In proliferative changes such as arterial intimal proliferation, proliferative glomerulonephritis, and interstitial cell infiltration, endothelial cells do not proliferate. This routine staining technique of endothelial cells by anti-A antiserum provide information not obtainable with light microscopy.