Vaccination with streptococcal extracellular cysteine protease (interleukin-1β convertase) protects mice against challenge with heterologous group A streptococci

Virtually all clinical isolates of group A streptococci secrete a highly conserved extracellular cysteine protease that cleaves human fibronectin and vitronectin, and converts IL-1β precursor to biologically active IL-1β. Based on the high degree of gene conservation within the species and its role in host pathogenicity, it was postulated that antibodies to the cysteine protease would confer protective immunity against S. pyogenes infection. To test this hypothesis, Swiss CD1 mice were intraperitoneally administered either saline, rabbit IgG, or IgG from rabbits immunized with the protease, and challenged with a highly virulent (minimum lethal dose 10 cfu) clinical isolate of S. pyogenes expressing a heterologous cysteine protease. The results indicate that mice administered IgG from rabbits immunized with purified cysteine protease had significantly enhanced survival when compared with mice given either non-specific rabbit IgG (log rank test; χ²; p = 0.0195) or saline (log rank test; χ²; p = 0.0002). Moreover, mice actively immunized with the cysteine protease had a significantly longer time to death than the control group (log rank test; χ²; p = 0.0418). The results show that the cysteine protease elicits non-type-specific immunity to challenge with heterologous S. pyogenes.     

Plant virus transmission by plasmodiophorid fungi is associated with distinctive transmembrane regions of virus-encoded proteins

Summary.  Computer analysis of published sequence data has consistently identified two complementary transmembrane domains in the coat protein readthrough domains of benyviruses, furoviruses and pomoviruses and in the P2 proteins of bymoviruses. These viruses differ in genome organisation but are all transmitted by plasmodiophorid fungi. The second domain is absent or disrupted in naturally-occurring deletion mutants that cannot be fungally-transmitted. In a non-transmissible substitution mutant of Beet necrotic yellow vein virus [Tamada et al. (1996) J Gen Virol 77: 1359–1367], the alignment of the helices is disrupted. From conserved patterns detected in transmembrane helix sequences and calculated relative helix tilts, structural arrangements consistent with tight packing of transmembrane helices were identified. These included ridge/groove arrangements between the two helices and strong electrostatic associations at the interfacial regions of the membrane. The data strongly suggest that these transmembrane helices facilitate the movement of virus particles across the fungal membrane. Accepted November 9, 2000 Received June 30, 2000     

Gastrointestinal graft-versus-host disease in recipients of autologous hematopoietic stem cells: incidence, risk factors, and outcome.

Graft-versus-host disease (GVHD) is seen in skin, intestinal mucosa, and liver after autologous stem cell transplantation. We reviewed 681 consecutive patients to estimate the probability of gastrointestinal (GI) GVHD, response to treatment, risk factors for development, and effect on survival. GI GVHD was defined by persistent symptoms, mucosal abnormalities at endoscopy, and histology showing apoptotic crypt cells with or without lymphoid infiltrates. The proportion of patients with GI GVHD was 90/681 (13%). Nausea and vomiting occurred in 90% and diarrhea in 40%. The mean time to developing symptoms was day +15, that to histologically proven diagnosis was day +42, and that to starting prednisone treatment was day +45 after stem cell infusion. Treatment with a short course of prednisone effected durable responses in 79% of patients, and an additional 18% responded to a second course of prednisone. A multivariable logistic regression model demonstrated that the combined factor of a diagnosis of breast cancer or hematologic malignancy and female sex was statistically significantly associated with the probability of GI GVHD (P = .003). Survival in patients with GI GVHD was not statistically different than that in those without GVHD. We conclude that women with breast cancer or hematologic malignancy are more likely to develop GI GVHD after autologous transplantation, and that treatment with prednisone was effective.     

The Effects of Discrimination Training on Voluntary Control of Cephalic Vasomotor Activity

The present study was conducted to test Brener's calibration theory of visceral learning. It was hypothesized that training in discrimination of cephalic vasomotor responses would enhance acquisition of voluntary and biofeedback control of cephalic vasomotor response. Four groups were given discrimination training to detect either constriction, dilation, both constriction and dilation, or given false feedback. All groups were then given biofeedback training in constriction. All groups were assessed on discrimination accuracy, acquisition of biofeedback control of constriction, and voluntary control of dilation and constriction. The results indicated that discrimination of cephalic vasomotor responses can be learned and that this skill facilitates the acquisition of biofeedback and voluntary control of vasomotor responses. This facilitation occurs only if discrimination training is given for the specific response to be learned. The data suggest that discrimination of the relevant response is a necessary but not sufficient condition for acquisition of voluntary control in biofeedback learning.     

Immuno- and genetic therapy in autoimmune diseases

Animal models of autoimmune disease have been developed that mimic some aspects of the pathophysiology of human disease. These models have increased our understanding of possible mechanisms of pathogenesis at the molecular and cellular level and have been important in the testing, development and validation of new immunotherapies. The susceptibility to develop disease in the majority of these models is polygenic as is the case in humans. The exceptions to this rule are gene knock outs and transgenic models of particular genes which, in particular genetic backgrounds, have also contributed to the understanding of single gene function and their possible contribution to pathogenesis. Gene therapy approaches that target immune functions are being developed with encouraging results, despite the polygenic nature of these diseases. Basically this novel immuno-genetic therapy harnesses the knowledge of immunology with the myriad of biotechnological breakthroughs in vector design and delivery. Autoimmune disease is the result of genetic dysregulation which could be controlled by gene therapy. Here we summarize the genetic basis of these human diseases as well as some of the best characterized murine models. We discuss the strategies for their treatment using immuno- and gene therapy.     

Glycoproteins present in human follicular fluid that inhibit the zona- binding capacity of spermatozoa

Previous studies have suggested that human follicular fluid contains factors that reduce the zona-binding capacity of spermatozoa. The present study provides further evidence of the existence of such factors. Using the hemizona binding assay (HZA), we have shown that the inhibitory effect of human follicular fluid on the zona-binding capacity of spermatozoa is concentration-dependent, an inhibitory effect being detected when the concentration of human follicular fluid was > or = 10%. A 1% concentration of human follicular fluid did not possess this inhibitory activity. Heating human follicular fluid at 56 degrees C for 30 min did not affect its inhibitory properties; treatment with proteinase-K abolished such inhibition. Human follicular fluid was fractionated sequentially by concanavalin-A affinity chromatography, Mono Q ion-exchange chromatography and Superose-12 gel filtration. The zona binding inhibitory activity resided in the fraction which bound to the lectin and Mono Q column and contained molecules with native molecular weights of 32 and 192 kDa. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis suggested that the 192 kDa glycoprotein was a tetramer, while the 32 kDa glycoprotein remained as a single molecular species under denaturing conditions. We conclude that two glycoproteins were responsible for the zona binding inhibitory activity of human follicular fluid. The physiological role of these factors remains unclear.      

Problems affecting performance of the fluorescent treponemal antibody-absorption test for syphilis.

Immunofluorescent staining of Treponema pallidum was studied to clarify the effect of three factors on the results of the fluorescent treponemal antibody-absorption test: (i) heat inactivation of sera at 56 degrees C for 30 min before testing, (ii) use of multicircle slides, and (iii) tungsten illumination to visualize and assess unstained treponemes on reactive as well as nonreactive smears. It was found that serum inactivation before testing was not necessary for detection of immunoglobin G antibody, but an immunoglobulin M prozone was detected in unheated serum. On multicircle slides, it was demonstrated that a false-positive reaction could be obtained in 30 s at 37 and 25 degrees C if a smear where a nonreactive serum had been placed was crossed by a strongly reactive serum from another circle. Tungsten illumination proved necessary for correct assessment of unstained treponemes on all fluorescent treponemal antibody-aborption test smears, reactive or nonreactive. The possible role of these factors in incorrect fluorescent treponemal antibody-absorption test results is discussed.     

Serum relaxin and the major endometrial secretory proteins in in-vitro fertilization and down-regulated hormone-supported and natural cycle frozen embryo transfer

Relaxin has been postulated to be a modulator of the expressionof the endometrial secretory proteins, insulin-like growth factorbinding protein (IGFBP-1) and placental protein 14(PP14). Thisstudy evaluated the expression of relaxin in relation to concentrationsof these secretory proteins along with oestradiol, progesteroneand human chorionic gonadotrophin in groups of pregnant andnon-pregnant patients who underwent differing assisted conceptiontreatments. Serum samples were taken from 88 patients at 8 and12 days after embryo transfer. At 12 days after embryo transfer,relaxin concentrations in the pregnant patients who had undergonein-vitro fertilization (IVF) or natural cycle frozen embryotransfer were significantly higher than those who did not conceivein these groups (mean concentrations 8334 versus 28 and 2608versus 62 pg/ml respectively, P <0.001). However concentrationsin the pregnant patients who had hormone support and transferof frozen embryos were not significantly different from thepatients who did not conceive after the same treatment. Althoughrelaxin expression was associated with corpus luteum activity,it was not related to the number of corpora lutea in IVF patients.A wide range of relaxin concentrations was seen to be compatiblewith a healthy pregnancy. These serum relaxin concentrationswere not found to be directly related to the serum concentrationsof IGFBP-1, PP14 or the other factors assessed in this study.     

Comparative studies and laboratory diagnosis of Vibrio vulnificus, an invasive Vibrio sp

Vibrio vulnificus was isolated from a bacteremic patient. This strain, together with other isolates of V. vulnificus, was compared with V. alginolyticus, V. fluvialis, and V. parahaemolyticus with regard to growth characteristics on enteric agar media (enabling isolation and identification) and production of exoenzymes which could correlate with invasive potential. V. vulnificus grew well on MacConkey. Endo, xylose-lysine deoxycholate, and Hektoen enteric agar plates. Because V. vulnificus colonies resembled those of lactose-fermenting strains of the family Enterobacteriaceae, however, isolation of this vibrio from mixed specimens or stools may require the use of thiosulfate-citrate-bile salts-sucrose agar. V. vulnificus produced numerous exoenzymes (protease, DNase, lipase, and esterase) but not elastase or lecithinase. Although differences in exoenzyme production were observed among the four vibrio species, no single exoenzyme could be linked to the invasive potential of V. vulnificus.