Peritoneal fluid concentrations of interleukin-8 in women with endometriosis: relationship to stage of disease

There is increasing evidence that immunological mechanisms play a role in the pathogenesis and pathophysiology of endometriosis. It was therefore of interest to study interleukin-8 (IL-8), a chemokine, in the peritoneal fluid and peripheral blood of women undergoing laparoscopic procedures. The presence and concentrations of IL-8 in relation to endometriosis, infertility and abdominal pain were evaluated. Samples of peritoneal fluid (n = 49) and peripheral blood (n = 50) were obtained from 50 consecutive patients undergoing laparoscopic surgery for various gynaecological indications (abdominal pain, infertility, sterilization). IL-8 was present in the peritoneal fluid of most women (87%). The concentration of IL-8 in the peritoneal fluid was higher in women with endometriosis compared to women without (P = 0.02). This difference was more pronounced in early (stage 1) endometriosis (P = 0.001). IL-8 concentrations in the peritoneal fluid were also higher in women with early endometriosis compared to women with later stages of the disease (P = 0.003). Peripheral blood concentrations did not correlate with peritoneal fluid concentrations of IL-8 and/or the presence of endometriosis. We conclude that IL-8 is an important factor that may contribute to the pathogenesis of endometriosis possibly by promoting neovascularization. This information can be a guide in the development of new therapeutic approaches for the treatment of endometriosis.      

Staphylococcal accessory regulator (sar) in conjunction with agr contributes to Staphylococcus aureus virulence in endophthalmitis.

Previous studies showed that an agr mutant strain of Staphylococcus aureus was partially attenuated in virulence compared to a parental strain in experimental endophthalmitis. The purpose of this study was to determine whether the sar locus, either alone or through interactions with agr, contributes to the regulation of virulence in S. aureus endophthalmitis. Experimental endophthalmitis was established by the midvitreous injection of approximately 30 CFU of S. aureus RN6390 or the isogenic attenuated strains RN6911 (agr mutant), ALC136 (sar mutant), and ALC135 (agr sar double mutant). Unexpectedly, the rate of reduction in electroretinographic B-wave amplitude in eyes infected with strain ALC136 (sar mutant) was not significantly different from the parental strain on postinfection day (PID) 5 (10% retention). In contrast, ALC135 (agr sar double mutant)-infected eyes retained 73% of preoperative B-wave amplitude on PID 5. Therefore, unlike agr, a mutation in the sar locus alone does not alter the overall virulence of wild-type S. aureus in experimental endophthalmitis. However, the combined effect of insertional mutations in both the sar and agr global regulators leads to near-complete attenuation of virulence.     

Expression of gtfS is essential for normal insoluble glucan synthesis by Streptococcus downei.

The gtfI and gtfS genes of Streptococcus downei were investigated to determine the contribution of the respective enzymes to glucan production in the presence and absence of other glucosyltransferases. Extracts of Escherichia coli expressing cloned gtfS produced a short linear dextran from sucrose which could act as a primer for insoluble glucan synthesis when mixed with extracts of a strain expressing recombinant gtfI. To elucidate the contribution of gtfS to glucan production by S. downei, a mutant was constructed by insertionally inactivating gtfS. S. downei (gtfS mutant) colonies exhibited a marked phenotypic change on sucrose-containing media and a decreased ability to adhere to glass and produced no detectable water-insoluble glucan. These experiments confirm that expression of gtfS is essential for normal insoluble glucan synthesis by S. downei.     

The induction of class I HLA by interferon-α is independent of the cell cycle,but the expression is enhanced by a G1/S block

The induction of class I HLA expression by interferon-α (IFN-α) was studied in lymphoid cells arrested or traversing different stages of the cell cycle. Exponential cultures of MOLT-4 cells and the MOLT-4 cell variant YHHH were treated with the cell cycle inhibitors aphidicolin and colcemid to obtain cell populations arrested in G₁/S and G₂/M, respectively, and also cells traversing from S to M and vice versa. Cytofluorimetry with the monoclonal antibody YTH/76.3 (which specifically detects those class I molecules which are most susceptible to IFN-α induction) was used to quantitate the class I HLA response to IFN-α. The results showed that the response to IFN-α is not restricted to a given stage of the cell cycle. These studies also revealed that when the cells were arrested at G₁/S, the absolute level of class I HLA expression was enhanced 2–3-fold, both in the presence or absence of either IFN-α or IFN-γ. Therefore, even when absolute levels changed, the ratio of IFN-induced expression to basal expression remained constant at all cell cycle stages. The level of expression of another surface antigen (the CD1 antigen HTA-1) was not affected by the G₁/S block. The results were confirmed by dot blot hybridization of poly (A)⁺ RNA using cDNA-specific probes. These findings suggest that the effect of IFN-α is continuous throughout the cell cycle but that a G₁-dependent event determines the extent of class I HLA expression, and leads to a synergistic superinduction by IFN in G₁/S-arrested cells.     

Multilamelled structures from the lungs of patients with pulmonary alveolar proteinosis

Multilamellated structures (MS), which accumulate in the lungs of patients with pulmonary alveolar proteinosis, were examined under the electron microscope. MS, in general, consisted of alternating light and dark lamellae arranged concentrically about membranous vesicles or electron-dense bodies. The darker osmiophilic lamellae consisted of trilaminar membranes 80 to 100 A thick, and the lighter lamellae, which varied from 150 to 300 A in width, appeared amorphous. MS made up 42.6 +/- 12.4% (n = 4) of the total particulate volume present in the pulmonary lavage effluents. MS resembled tubular myelin structures from the lungs of patients, although these latter structures accounted for only 1.6 +/- 2.3% (n = 4) of the total particulate volume. Abnormalities, such as polygonal tubules, irregular tubule dimensions, and large size potential, indicated that tubular myelin structures in the lungs of patients were variable in structure. MS appear to be a form of tubular myelin structure. MS were treated with a variety of disruptive agents and then examined under the electron microscope. Information was obtained about the composition of the MS and the intermolecular forces involved in their three-dimensional organization. Extraction of the MS with acetone or treatment of the structures with phospholipase c resulted in loss of membranes from the structures without a concomitant loss of the intermembranous amorphous material, indicating that phospholipids were a major membrane component. Analysis of phospholipids in the acetone extracts of the MS revealed that the membranes consisted of 45% phosphatidylcholine and that the major fatty acid of the phosphatidylcholine was palmitate (76%). The amorphous material of the intermembranous spaces consisted primarily of protein, since it was destroyed by the proteases trypsin and pronase without loss of the membranes. Reducing agents such as mercaptoethanol and dithiothreitol were disruptive, indicating the importance of disulfide bridges between the protein constituents in maintaining the integrity of the MS. The significance of hydrophobic interactions between the protein constituents was demonstrated by the disruptive action of chaotropic agents according to the sequence KSCN greater than KI greater than KCl. These data indicate that MS consist of phospholipid membranes organized in a protein matrix maintained by intermolecular disulfide bridges and hydrophobic interactions.     

Circulating immune complexes during immunotherapy.

Chronic immunization may lead to the production of circulating immune complexes (CICs). This study was undertaken to determine the presence of circulating IgG immune complexes in 95 subjects with allergic rhinitis/asthma receiving immunotherapy, 46 individuals with similar diagnosis but not on immunotherapy, and 64 healthy controls. Modified Raji cell and murine leukemia cell (L-1210) assays, selected for a high density of Fc receptors and devoid of Epstein-Barr virus membrane antigen, were used. Other immunological parameters such as immunologlobulins G, A, M, E, and rheumatoid factor activity were also studied. The CIC concentrations in the treated group did not differ significantly from the untreated group, although both groups did have a significantly higher concentration than the healthy controls. The presence of CICs has no relationship with age or sex of patients, dosage of allergen administered, number and nature of allergens received, period between the time of last injection and the blood sampling, and the duration of immunotherapy. Serum IgG, IgA, IgM, and rheumatoid factor activity did not differ between the treated and untreated groups. IgE was significantly higher in the treated group when compared with the untreated, and IgE levels in treated patients with elevated CICs were significantly increased compared with CIC-negative treated patients. These data suggest that CICs are present in serum of atopic diseases such as allergic rhinitis/asthma. Significantly, an association of elevation of CICs with immunotherapy could not be demonstrated.