T HELPER 1/2 LYMPHOCYTE URINARY CYTOKINE PROFILES IN RESPONDING AND NONRESPONDING PATIENTS AFTER 1 AND 2 COURSES OF BACILLUS CALMETTE-GUERIN FOR SUPERFICIAL BLADDER CANCER

PURPOSE: Interleukin (IL)-2 and interferon-gamma are released during T helper 1 lymphocyte responses and IL-10 is released during T helper 2 lymphocyte responses. We have previously reported that a T helper 1 lymphocyte urinary cytokine profile is associated with a favorable prognosis after bacillus Calmette-Guerin (BCG) treatment. We evaluated the T helper 1/2 lymphocyte cytokine profiles during courses 1 and 2 of 6 weekly BCG instillations. MATERIALS AND METHODS: Urinary interferon-gamma, IL-2 and IL-10 were measured by enzyme-linked immunosorbent assay after each of 6 weekly instillations of 150 mg. BCG, Pasteur strain, in 19 patients with superficial stages Ta and T1 bladder cancer, and carcinoma in situ. The 11 patients who did not respond to course 1 were re-treated according to the same schedule and reevaluated. RESULTS: During course 1 interferon-gamma was higher than during course 2 (p <0.001), which was associated with nonrecurrence (p <0.001). In contrast, IL-2 cytokine was higher after course 2 (p <0.01), which was associated with a BCG response (p = 0.01). Interferon-gamma and IL-10 correlated during courses 1 and 2 (p = 0.04 and 0.0004, respectively). We distinguished groups 1-immediate T helper 1 lymphocyte profile consisting of responders to course 1 with high interferon-gamma, IL-2 and IL-10, 2-delayed T helper 1 lymphocyte profile consisting of responders to course 2 with early high IL-2 and 3-consisting of nonresponders to the 2 courses with low interferon-gamma, IL-2 and IL-10. CONCLUSIONS: A T helper 1 lymphocyte urinary cytokine profile was associated with a clinical response to BCG. A repeat BCG course induces a favorable immune response in a subset of patients, suggesting that maintenance therapy may be beneficial.     

ANTI-HIV AND -SIV IMMUNITY IN THE VAGINA

Most HIV infections worldwide are transmitted through heterosexual contact. In order to develop vaccination strategies, the basic biology of the immune system in female reproductive tract and the full range of vaginal immune responses that occur during natural HIV infection must be understood. The cervicovaginal mucosa contains a complete set of immune cells, including antigen-presenting cells, CD4+ and CD8+ T cells, and B cells. The CVS of HIV-infected women and SIV-infected female rhesus macaques contain variable levels of antiviral antibodies. Some of this variation is due to the effects of female ovarian hormone cycle. IgG antibodies make up the bulk of the antiviral antibody response. However, IgA antibodies are present at lower levels. HIV/SIV-specific CD8+ cytotoxic T lymphocytes are present in the cervicovaginal mucosa of infected women and rhesus macaques. A vaccine that can elicit strong antiviral immunity may provide protection for hetorosexual HIV-1 transmission.     

Interaction of mammalian sperm nuclear protamines and peptides derived thereof with immobilized zinc

The interaction of mammalian and human protamines with zinc was studied by immobilized metal ion affinity chromatography (IMAC). The affinity of protamines containing blocked cysteine residues was found to correlate in part with the presence and number of histidine residues in the protamine structure: absence or low affinity of P1 protamines containing 0 or 1 histidine residue; high affinity of human P2 protamine containing 9 histidines. Nevertheless a fraction strongly retained on an IDA-Zn(II) column was observed for P1 protamines with one histidine in the N-terminal sequence (ram and boar protamines). The strong binding was found to be related to the presence of tyrosine, serine and threonine closely spaced to the histidyl side chain. In the case of human protamine P2, the strong retention on the IDA-Zn(II) column seems to result from the additive contribution of all the histidine residues of the molecule. Thus, strong retention of protamines in IMAC seems to depend on an additive contribution of amino-acid side chains: histidine, tyrosine, serine, threonine and perhaps arginine. The high affinity of protamines, more especially P2 protamines, for zinc suggests that this metal ion could play a role for their correct folding and binding to DNA.     

Increased expression of CD11b/CD18 on phagocytes in ischaemic disease: a bridge between inflammation and coagulation

The aim of this study was to assess the expression of CD11b/CD18 integrin adhesion molecules on the phagocytes of patients with ischaemic diseases, and to evaluate the concentration of soluble adhesion molecules that are released from endothelium (sICAM-1) and from phagocytes (sL-selectin). A total of 370 patients were enrolled: 120 with coronary artery disease (CAD); 50 with peripheral artery occlusive disease (PAOD); and 200 control subjects with no clinical manifestations of ischaemic disease. CD11b/CD18 integrin was detected by flow cytometry, whereas sL-selectin and sICAM-1 concentrations were detected using a sandwich-type immunoassay. CD11b/CD18 integrin expression was found to be higher in the patients with ischaemic disease than in the control subjects ( P  < 0.001). The PAOD patients had higher values of CD11b/CD18 integrin than the CAD ones ( P  < 0.01). The concentration of soluble adhesion molecules did not show any significant differences within the three groups ( P  = NS). The high expression of CD11b/CD18 integrin in ischaemic disease patients may depend on the increased, but probably stable, cytokine network that has been demonstrated to occur in chronic ischaemic diseases: the difference observed between PAOD and CAD patients could be the consequence of higher inflammatory activation probably resulting from the greater extent of the atherosclerotic process in PAOD, or of the more localized ischaemic area in CAD patients. CD11b/CD18 can therefore be considered a marker of chronic phagocyte activation during ischaemic disease. On the other hand, sICAM and sL-selectin concentrations were found to be within the normal range; they have recently been considered as a marker for acute ischaemic events and acute inflammatory process activation. Our results confirm that in uncomplicated atherosclerosis no acute inflammatory process activation should occur.     

Mycobacterium tuberculosis chaperonin 10 and N-truncated fragments Their synthesis and purification by the isoelectric focusing technique carried out in solution

The Mycobacterium tuberculosis chaperonin 10 protein and fragments corresponding to sequences 59-99, 51-99 and 26-99 were synthesised by the solid-phase methodology using a double coupling protocol and without the aid of capping agents. After the final acid cleavage using the low TFMSA-high HF protocol the polypeptides were purified by either the ion exchange chromatography/RP-HPLC combination or the isoelectric separation carried out in solution and followed by semi-preparative RP-HPLC. Comparison of the results obtained through the two approaches indicated that in general the isoeletricfocusing/HPLC combination was superior both in terms of recovery of final material and its purity. The advantages found were as follows: (i) Unlike ion exchange chromatography, no tailoring of the separation conditions is required. (ii) Several consecutive focusings can be carried out in progressively narrower pH gradients. This increases the separation resolution without the need of changing other separation parameters, (iii) Very little manipulation is needed, and each focusing requires 3-5 h. (iv) Full compatibility with non-ionic denaturants such as 8 M urea. This increases solubility so that using the ROTOFOR instrument described here 50-100 mg crude polypeptide can be processed daily. Thus the isoelectric focusing technique carried out in solution is a valid and inexpensive alternative to ion exchange chromatography. © Munksgaard 1997.     

Upgrading from Single Chamber Right Ventricular to Biventricular Pacing in Permanently Paced Patients with Worsening Heart Failure: The RD-CHF Study

Background: Biventricular (BiV) stimulation lowers morbidity and mortality in patients with drug-refractory congestive heart failure (CHF), depressed left ventricular (LV) function, and ventricular dyssynchrony in absence of indication for permanent cardiac pacing. This pilot, single-blind, randomized, cross-over study examined the safety and efficacy of upgrading conventional pacing systems to BiV stimulation in patients with advanced CHF .
Methods: We included 56 patients in New York Heart Association (NYHA) functional classes III or IV despite optimal drug treatment and ventricular dyssynchrony (interventriclar delay >40 ms or LV preejection delay >140 ms) in need of pacemaker replacement. We compared the patients' functional status, arrhythmias, and standard echocardiographic measurements during 3 months of conventional, single right ventricular (RV) versus 3 months of BiV stimulation .
Results: There were 44 patients in the cross-over phase. QRS duration was shortened by 23% and LV preejection delay by 16% with BiV stimulation. NYHA functional class, 6-minute hall walk and quality of life score were significantly improved with BiV stimulation compared with single RV pacing by 18%, 29%, and 19%, respectively. No significant difference was observed in the ventricular arrhythmia burden or LV reverse remodeling between the 2 periods .
Conclusions: This pilot study showed that upgrading from RV pacing to BiV pacing significantly improves symptoms and exercise tolerance in chronically paced patients with advanced CHF and mechanical dyssynchrony .