Antitubercular antibodies to antigens of various molecular weights in patients with active pulmonary tuberculosis

Antibodies were determined in 72 tuberculous patients and 27 healthy donors using enzyme immunoassay (EIA) and immunoblotting (IB). It was shown that the levels of the antibodies to BCG and H37Rv sonicates were lower in the patients with disseminated tuberculosis than in the patients with fibrocavernous or infiltrative tuberculosis. Significant differences in the antibody spectra in the patients with different forms of tuberculosis were also shown. Thus, in the patients with disseminated tuberculosis IB with BCG and H37Rv sonicates revealed only single bands in the region of the reaction with the antigens of low molecular weights. In the patients with infiltrative tuberculosis the reaction with H37Rv or BCG sonicates most frequently revealed the antibodies to the antigen determinants with molecular weights of 50, 30, 41, 54, 52 and 60.43 or 45.5, 40.5, 57 and 52 kD, respectively. In the patients with fibrocavernous tuberculosis the reaction with H37Rv or BCG sonicates most frequently revealed the antibodies to the antigen determinants with molecular weights of 54, 42, 48, 30 and 13.5 or 24, 57, 37 and 14.5 kD respectively.     

Human Rh D monoclonal antibodies (BRAD-3 and BRAD-5) cause accelerated clearance of Rh D+ red blood cells and suppression of Rh D immunization in Rh D- volunteers

The use of prophylactic anti-D to prevent Rh D immunization in Rh D- women and subsequent hemolytic disease in Rh D+ infants is widespread, but has led to shortages of the anti-D Ig. With the aim of substituting monoclonal anti-D for Rh D prophylaxis, we have compared the abilities of monoclonal and polyclonal anti-D to clear Rh D+ red blood cells (RBCs) infused into Rh D- male volunteers and to suppress Rh D immunization. Two human monoclonal antibodies (MoAbs), BRAD-3 (IgG3) and BRAD-5 (IgG1), produced from stable Epstein-Barr virus-transformed B-lymphoblastoid cell lines, were selected because of their proven in vitro activity in promoting RBC lysis in antibody-dependent cell- mediated cytotoxicity assays. RBC clearance was assessed by intravenous injection of 3 mL of 51chromium-labeled D+ RBCs into 27 volunteers 48 hours after intramuscular injection of monoclonal or polyclonal anti-D. Further 3-mL injections of unlabeled D+ cells were administered at 6 and 9 months to induce immunization. Blood samples were taken throughout the 12-month period of study for the serologic detection of anti-D. The mean half-life (t50%) of RBCs in 7 recipients of 300 micrograms BRAD-5 (5.9 hours) was similar to that in 8 recipients of 500 IU polyclonal anti-D (5.0 hours), whereas D+ cells were cleared more slowly in some of the 8 subjects injected with 300 micrograms BRAD- 3 (mean t50% 12.7 hours) and in 1 individual administered 100 micrograms BRAD-3 (t50% 41.0 hours). The rate of RBC clearance in both groups administered 300 micrograms monoclonal anti-D correlated with the amount of antibody bound per cell, determined by flow cytometry. There was no evidence of primary immunization having occurred in any subject after 6 months of follow-up. Five of 24 subjects produced anti- D after one or two further injections of RBCs, confirming that they were responders who had been protected by the monoclonal or polyclonal anti-D administered initially. Four of these responders were recipients of monoclonal anti-D (3 BRAD-3, 1 BRAD-5). One individual who received BRAD-5 produced accelerated clearance of D+ RBCs at the third unprotected RBC challenge but did not seroconvert. This study shows that the human MoAbs BRAD-3 and BRAD-5 can prevent Rh D immunization, and indicates that they may be suitable replacements for the polyclonal anti-D presently used in prophylaxis of Rh D hemolytic disease of the newborn.(ABSTRACT TRUNCATED AT 400 WORDS)     

The L4F3 antigen is expressed by unipotent and multipotent colony- forming cells but not by their precursors

Antibody L4F3 is a murine monoclonal antibody that recognizes an antigen expressed on in vitro colony-forming cells, including virtually all CFU-GM, CFU-Meg, BFU-E, and CFU-Mix. In the present study we examined whether cells that do not express the L4F3 antigen include precursors of hematopoietic colony-forming cells. Colony-forming cells were depleted from marrow by treatment with L4F3 and complement. The remaining cells generated CFU-GM, BFU-E, and CFU-Mix when cultured in the presence of irradiated adherent cell layers from long-term marrow cultures. Marrow cells not expressing the L4F3 antigen, which were separated by cell-sorting techniques, were depleted of colony-forming cells but nevertheless generated CFU-GM when cultured over irradiated adherent cell layers. These data suggest that there are marrow precursors that do not express the L4F3 antigen and that give rise to colony-forming cells of multiple types. Negative selection techniques should allow the enrichment of these precursors of colony-forming cells, thereby enabling direct studies of these immature stem cells.     

Monoclonal antibody 12-8 recognizes a 115-kd molecule present on both unipotent and multipotent hematopoietic colony-forming cells and their precursors

A monoclonal antibody, 12-8, prepared against KG-1a cells, recognizes an approximately 115-kd cell surface antigen and reacts with 3% to 4% of bone marrow cells, including most of the blast cells. The antigen is not expressed on peripheral blood cells. Marrow cells expressing 12-8 collected by fluorescence-activated cell sorting contained nearly all of the unipotent (CFU-GM, BFU-E) and multipotent (CFU-MIX) colony- forming cells. The isolated 12-8 positive marrow population also contained precursors of these colony-forming cells. In a two-stage long- term marrow culture system employing irradiated allogeneic marrow adherent cells, 12-8 positive cells produced both unipotent and multipotent colony-forming cells for ten weeks. Moreover, the output of colony forming cells substantially exceeded the input. Antibody 12-8 appears useful for analysis and possibly enrichment of hematopoietic progenitor cells that include colony-forming cell precursors.     

Genetic engineering production of H37Rv M. tuberculosis proteins]

A complete library of M. tuberculosis H37Rv genes was produced by incorporating the DNA fragments of M. tuberculosis H37Rv into the lambda pSI phasmid. For this, DNA isolated from the mycobacteria was treated by EcoRI restrictases and the fragments of 8-17 thousand nucleotide pairs were crosslinked with the phasmid DNA. Hybrid DNA molecules were packed into the caspids from the proteins of E. coli BHB2688 and BHB2690 strains. By estimates, this library contained 98% of M. tuberculosis H37Rv genome so that any required gene can be found at 0.99 probability. The needed genes were sought by monoclonal antibodies against a protein with a molecular mass of 17-19 kDa (IT-12, IT-51, IT-54) obtained from the WHO. The protein gene was also produced by the method for raising the end sequences using synthesis of two oligonucleotides SP30 complementary to segments that limit this gene in the presence of Taq-DNA-polymerase and DNA of M. tuberculosis H37Rv. Copies of a gene (MT-1) were produced by denaturation, firing and raising. This gene was incorporated in the puC119 plasmid and expressed in E. coli cells, the direction of reading being checked up. A protein with a molecular mass of 19 kDa was detected in E. coli extracts with the expressed pMT-2 plasmid using monoclonal antibodies IT-12 and IT-54 in enzyme-linked immunoassay and immunoblotting.     

Maximum Tolerated Osmolarity for Peripheral Administration of Parenteral Nutrition in Pediatric Patients

Background: Limited data support a recommended maximum osmolarity for administration of peripheral parenteral nutrition (PPN). In this retrospective, matched‐cohort study, we evaluated the incidence of phlebitis or infiltration associated with administration of PPN with an osmolarity >1000 mOsm/L vs ≤1000 mOsm/L. Materials and Methods: Patients ≤18 years old who received PPN in a 2‐year period were included in the study. Data related to patient demographics, PPN constituents, and adverse effects were analyzed. Results: A total of 352 patients met entry criteria. Overall, 139 (40%) patients experienced phlebitis or infiltration. There were no differences between patients who did or did not develop adverse events in terms of age or weight. Administration of PPN with osmolarity >1000 mOsm/L vs ≤1000 mOsm/L significantly increased infiltration (17% vs 7%; odds ratio [OR, 2.47]; 95% confidence interval [CI], 1.24–4.94; P = .01) and the combined composite end point of phlebitis or infiltration (45% vs 34%; OR, 1.65; 95% CI, 1.07–2.54; P = .02). In multivariate analysis, osmolarity >1000 mOsm/L vs ≤1000 mOsm/L was an independent risk factor for developing complications (OR, 1.67; 95% CI, 1.08–2.52; P = .02). Conclusion: Two of every 5 children experienced phlebitis or infiltration during administration of PPN. These adverse effects were more often observed in those who received PPN with osmolarity >1000 mOsm/L vs ≤1000 mOsm/L. With this high incidence of adverse effects, we recommend that if PPN is used, the osmolarity should not exceed 1000 mOsm/L. More important, PPN should only be used temporarily until central access is obtained.     

The hypophyseo-thyroid system in psoriatic arthritis combined with chronic opisthorchiasis

Paper presents the data on the content of hypophysial-thyroid and hypophysial-adrenal systems' hormones obtained by means of radioimmune assay, and the data of timecourse hepatoscintibiligraphy with Tc 99 m TCK-15 of 94 patients (first group--48 patients with psoriatic arthiritis combined with chronic opisthorchiasis; second group--46 pure psoriatic arthritis patients). Statistically significant increase in the content of triiodinethyronine, thyroxine, hypophysial thyreotropic hormone, corticotropic hormone and significant reduction of cortizol and aldosteron contents were observed in case of mixed pathology. Thyroid hormones' level was directly connected with hepatic and gallbladder disfunctions. The above-mentioned changes belong to pathogenetic mixed-pathology factors which should be taken into account in elaboration of best treatment methods.