C1q,a subunit of the first component of complement,enhances antibody-mediated apoptosis of cultured rat glomerular mesangial cells

We have shown previously that IgG2a anti-Thy-1 MoAb (ER4G) induces apoptosis of rat mesangial cells (GMC) in vitro. Since the classical complement pathway plays an essential role in Thy-1 nephritis, we analysed whether C1q, a subunit of the first component of complement, enhances the ER4G-mediated apoptosis of rat GMC. Two different subclasses of anti-Thy-1 MoAb, ER4G (IgG2a) and ER14 (IgG1), were used. It was established that ER4G binds C1q efficiently, while ER14 reacts poorly with C1q. For the experiments of apoptosis, quiescent rat GMC were exposed for 1 h at 37°C to a fixed concentration of anti-Thy-1 MoAb and incubated further for 16 h at 37°C in the presence or absence of C1q. GMC exposed to medium (M-GMC) followed by incubation of the cells with medium alone was used as controls. Apoptosis was assessed by morphological studies and quantitative analysis on FACS using FITC-annexin V (the annexin V methods) or bicolour FACS analysis using FITC-annexin V and propidium iodide (the annexin V/PI method). With the annexin V method, M-GMC revealed 9.4 ± 1.4% apoptosis. C1q had only marginal effects on apoptosis of M-GMC. GMC exposed to ER4G (ER4G-GMC) and further incubated with medium in the absence of C1q resulted in 25.7 ± 5.7% apoptosis (P < 0.01 relative to control). Incubation of ER4G-GMC together with 100 μg/ml of C1q significantly increased GMC-apoptosis up to 39.4 ± 4.9% (P < 0.01 relative to ER4G-GMC incubated in the absence of C1q). This enhancing effect of C1q on apoptosis of ER4G-GMC was time- and dose-dependent. In contrast, C1q did not significantly alter the apoptosis of either GMC exposed to ER14 (ER14-GMC) or to F(ab′)₂-ER4G (F(ab′)₂-ER4G-GMC), while ER14-GMC or F(ab′)₂-ER4G-GMC incubated with medium resulted in significant apoptosis compared with control. These results were supported by morphological studies and bicolour FACS analyses in time course experiments using the annexin V/PI method. The effect of C1q is dependent on the presence of intact C1q-containing globular heads and does not occur with collagen-like fragments of C1q. Furthermore, incubation of ER4G-GMC with anti-mouse κ-chain antibodies also increased ER4G-mediated GMC-apoptosis. These results indicate for the first time that C1q enhances antibody-mediated apoptosis of rat GMC in vitro, presumably by its binding to ER4G and probably by additional cross-linking of Thy-1 on the surface of GMC.     

Direct oral questions to blood donors: the impact on screening for human immunodeficiency virus

BACKGROUND: In December 1990, the Food and Drug Administration recommended that all United States blood centers implement a policy of asking prospective donors direct oral questions (DOQs) about human immunodeficiency virus (HIV) risk behaviors to increase the safety of the blood supply. STUDY DESIGN AND METHODS: To evaluate the impact of the DOQ policy, HIV-related deferral and HIV seroprevalence data were analyzed at four American Red Cross blood centers for the year before the policy change and the year after. An epidemiologic analysis with stratification was conducted, including the calculation of odds ratios (OR) and 95-percent CIs. RESULTS: Two of the four blood centers showed an overall significant increase in HIV-related deferral after implementation of the DOQ policy: OR = 4.04, (95% CI = 3.41, 4.76); OR = 2.93, (95% CI = 2.67, 3.21). The increase in HIV-related deferral was higher for women. HIV seroprevalence decreased at all four centers, including the two that did not see an increase in HIV-related deferrals. Seroprevalence declined by 14 percent in the two centers with increases in HIV-related deferral, which was neither significant nor attributable to DOQs. CONCLUSION: Given that HIV antibody screening cannot detect HIV-seronegative (but infectious) "window-period" donations, the deferral of at-risk donors may offer some additional protection to the blood supply. However, evidence was not found of an increase in safety of the blood supply as measured by HIV seroprevalence.     

T (Thomsen–Friedenreich) antigen and other simple mucin-type carbohydrate antigens in precursor lesions of gastric carcinoma

In a previous report we suggested that T antigen appeared to be associated with gastric carcinoma. To verify this hypothesis and characterize the pattern of expression of simple-mucin type carbohydrate antigens (Tn. sialyl-Tn and T before and after neuraminidase) in normal gastric mucosa and precursor lesions of gastric carcinoma, we studied the mucosa adjacent to 100 cases of gastric carcinoma, gastric biopsies of 60 dyspeptic patients, eight adenomatous polyps and eight hyperplastic polyps. The expression of the antigens was more related to the cell type and underlying lesions than to the coexistence of carcinoma. The most distinctive findings concerned intestinal metaplasia, dysplasia and hyperplastic lesions. In intestinal metaplasia, Tn was found mostly in columnar cells and sialyl-Tn in goblet cells. T was more prevalent in incomplete intestinal metaplasia than in complete. A high prevalence of sialyl-Tn expression and cell membrane immunoreactivity for T antigen, similar to those previously found in gastric carcinomas, were observed in three adenomatous polyps, one hyperplastic polyp, five cases of adenomatous dysplasia in the neighbourhood of intestinal carcinomas and four cases of marked foveolar hyperplasia, three of which were from the mucosa adjacent to diffuse carcinomas. We conclude that adenomatous and hyperplastic lesions share with gastric carcinomas features of aberrant glycosylation, namely the cell membrane expression of T antigen.     

Deposition of IgA is Associated with Macrophage Influx in the Kidney of Rats

In the present study we treated rats with N-nitrosodimethylamine (DMN) or D-galactosamine (GALN) to achieve increased circulating IgA levels in rats. GALN-treated rats showed a six-fold increase in serum IgA levels after the first intraperitoneal (i.p.) injection, whereas a 10-fold increase after a second i.p. injection of GALN was seen. DMN-treated rats showed a three-fold increase in serum IgA levels. No differences were observed in IgG and IgM levels between treated and non-treated rats. Sequential renal biopsies analysed by immunofluroescence exhibited mesangial deposits of IgA with different intensities of C3 deposition. Rats treated with GALN showed more IgA deposition in the kidney than DMN-treated rats. The IgA deposition together with C3 was more prominent in rats treated with GALN than in rats treated with DMN. The deposition of C3 together with IgA was associated with an influx of monocytes as detected by ED-1, an antibody directed against a rat monocyte marker. These studies provide evidence that an increase in serum IgA levels is associated with deposition of IgA in the kidney and that IgA has an inflammatory potential.     

Development of a Patient Education Program for New Amputees

In this article, the authors describe how the needs of the patient with a new amputation were assessed and how behavioral objectives were developed for an educational program that would address the knowledge, motor skills, and attitudes necessary for self-care. Two teaching techniques are described. An evaluation process that identifies the patient's or family's achievement of the objectives is discussed.