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Biomedical subjects

G Russ

Publications and source records attributed to G Russ.

At least 55 records · Page 3Linked to original sources

Role of pulmonary blood flow in postpneumonectomy lung growth.

To study the influence of blood flow on postpneumonectomy lung growth, we banded the left caudal lobe pulmonary artery of eight ferrets in such a way that blood flow to the caudal lobe did not increase when the right lung was excised 1 wk later. The fraction of the cardiac output received by the right lung before pneumonectomy was therefore directed entirely to the left cranial lobe. Three weeks after pneumonectomy the weight, volume, and protein and DNA contents of the two lobes of the left lung were measured and compared with those of five unoperated animals and eight animals after right pneumonectomy alone. Although its perfusion did not increase after pneumonectomy, the left caudal lobe of banded animals participated in compensatory growth, increasing in weight and protein and DNA contents. Although the cranial lobe of banded animals received 25% more of the cardiac output than the same lobe in pneumonectomized animals, cranial lobe volume and protein and DNA contents in the two groups were similar. Caudal lobes were smaller in banded than in simple pneumonectomized animals and tended to contain less protein, whereas the cranial lobes tended to be heavier. We conclude that increased pulmonary perfusion is not necessary for compensatory lung growth in adult ferrets, but it may modify this response.

Animals↗

Reduced expression of HLA-DP antigens on PWM stimulated T lymphocytes in patients suffering from psoriasis vulgaris.

24 psoriasis vulgaris patients were investigated for the expression of class II HLA antigens on the surface of PWM-stimulated T lymphocytes. The percentage of the expression of HLA-DP antigens ranged from 50.1% to 82.6% compared to a 100% level in healthy controls (p less than 0.005). No significant differences in the expression of HLA-DR antigens were observed. A higher frequency of some HLA antigens was found in the group of patients studied: B13 - 23.1%/6.2%, B 17 - 15.4%/7.1%, and Dw7 - 59.4%/15.8%.

Concanavalin A↗

Influenza virus hemagglutinin trimers and monomers maintain distinct biochemical modifications and intracellular distribution in brefeldin A-treated cells.

Brefeldin A (BFA) induces the retrograde transport of proteins from the Golgi complex (GC) to the endoplasmic reticulum (ER). It is uncertain, however, whether the drug completely merges the ER with post-ER compartments, or whether some of their elements remain physically and functionally distinct. We investigated this question by the use of monoclonal antibodies specific for monomers and trimers of the influenza virus hemagglutinin (HA). In untreated influenza virus-infected cells, monomers and trimers almost exclusively partition into the ER and GC, respectively. In BFA-treated cells, both monomers and trimers are detected in the ER by immunofluorescence. Cell fractionation experiments indicate, however, that whereas HA monomers synthesized in the presence of BFA reside predominantly in vesicles with a characteristic density of the ER, HA trimers are primarily located in lighter vesicles characteristic of post-ER compartments. Biochemical experiments confirm that in BFA-treated cells, trimers are more extensively modified than monomers by GC-associated enzymes. Additional immunofluorescence experiments reveal that in BFA-treated cells, HA monomers can exist in an ER subcompartment less accessible to trimers and, conversely, that trimers are present in a vesicular compartment less accessible to monomers. These findings favor the existence of a post-ER compartment for which communication with the ER is maintained in the presence of BFA and suggest that trimers cycle between this compartment and the ER, but have access to only a portion of the ER.

Animals↗

Type-common and type-specific monoclonal antibodies to herpes simplex virus types 1 and 2.

Seventeen monoclonal antibodies (Mabs) reacting specifically with the cells infected with herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) were characterized by a variety of immunological tests such as radioimmunoprecipitation, immunoblotting and virus-neutralization. The majority of Mabs was directed against glycoprotein B (anti-gB), six reacted with glycoprotein C (anti-gC) and one with glycoprotein G (anti-gG). Six anti-gB Mabs reacted with both types of HSV (anti-gB-1,2), two anti-gB and all the six anti-gC Mabs have been specific for HSV-1 (anti-gB-1 and anti-gC-1). The remaining two anti-gB Mabs and the anti-gG have been specific to HSV-2 (anti-gB-2 and anti-gG). Only three out of the seventeen examined Mabs neutralized the virus.

Animals↗

[The effect of certain substances on changes in the expression of class II HLA antigens].

With the aid of the RIA method, the authors found that the stimulation of T-lymphocytes by means of PWM lectin is linked to the expressivity of HLA antigens of class II on their surface. A simultaneous addition of indomethacin, tetracycline, coreton, hydrocortisone and novozir was decreasing the expressivity of the Ia antigens and, at the same time inhibited the lymphocyte proliferation. The growth factor for T (IL-2) lymphocytes was increasing the expressivity of HLA-DR antigens in the cells even without a previous stimulation by mitogen. Glucan, serous thymic factor, natrium salicylicum, hippuric acid had no effect on the expressivity of the HLA antigens of class II.

HLA-D Antigens↗

Solitary increased tibial uptake of 99mTc-diphosphonate unmasking pancreatic tumor-related medullary fat necrosis.

Pancreatic inflammation and tumors can induce various systemic lesions of steatonecrosis. We report here the case of a 73-year-old woman presenting a painful left leg. Roentgenograms and tomograms of the left tibia were normal. Radionuclide bone scan showed diffuse increased uptake in the whole tibia and a CT scan of the same region demonstrated an unusual pattern of bone tumor. Tibial biopsy revealed intra medullary steatonecrosis and led to the discovery of a pancreatic carcinoma.

Adenocarcinoma↗

Brefeldin A redistributes resident and itinerant Golgi proteins to the endoplasmic reticulum.

Brefeldin A (BFA) has been reported to block protein transport from the ER and cause disassembly of the Golgi complex. We have examined the effects of BFA on the transport and processing of the vesicular stomatitis virus G protein, a model integral membrane protein. Delivery of G protein to the cell surface was reversibly blocked by 6 micrograms/ml BFA. Pulse-label experiments revealed that in the presence of BFA, G protein became completely resistant to endoglycosidase H digestion. Addition of sialic acid, a trans-Golgi event, was not observed. Despite processing by cis- and medial Golgi enzymes, G protein was localized by indirect immunofluorescence to a reticular distribution characteristic of the ER. By preventing transport of G protein from the ER with the metabolic inhibitor carbonyl cyanide m-chlorophenylhydrazone or by use of the temperature-sensitive mutant ts045, which is restricted to the ER at 40 degrees C, we showed that processing of G protein occurred in the ER and was not due to retention of newly synthesized Golgi enzymes. Rather, redistribution of preexisting cis and medial Golgi enzymes to the ER occurred as soon as 2.5 min after addition of BFA, and was complete by 10-15 min. Delivery of Golgi enzymes to the ER was energy dependent and occurred only at temperatures greater than or equal to 20 degrees C. BFA also induced retrograde transport of G protein from the medial Golgi to the ER. Golgi enzymes were completely recovered from the ER 10 min after removal of BFA. These findings demonstrate that BFA induces retrograde transport of both resident and itinerant Golgi proteins to the ER in a fully reversible manner.

Animals↗

Inhibition of influenza virus haemolytic and haemagglutination activities by monoclonal antibodies to haemagglutinin glycopolypeptides HA1 and HA2.

Acid treatment of influenza virus enhanced haemagglutination inhibiting (HI) activity of some anti-HA1 monoclonal antibodies (MoAbs). These changes in the HI-activity could be either due to alteration in the mutual orientation of MoAb (e.g. IC8, IB8) binding epitope to receptor site or to an increase in the number of epitopes accessible to the corresponding MoAbs (e.g. IVA1). HI test with pH 5-virus revealed similar (although not identical) antigenic differences among related virus strains as the HI test with pH 7-virus. Anti-HA2 MoAbs were negative in the HI test with both pH 5- and pH 7-virus. Anti-HA1 MoAbs showed a HI activity with pH 5-treated BHA similar to that with pH 5-treated virus. Surprisingly one out of eight anti-HA2 MoAbs (IIF4) exhibited a relatively high HI activity to pH 5-BHA-mediated haemagglutination. Virus-induced red blood cell haemolysis was efficiently inhibited with several anti-HA1 MoAbs (e.g. IC8, IB8, and IIB4) while other anti-HA1 antibodies, including IVA1 and IVG6 with preferential reactivity with pH 5-treated antigens in RIA, gave no inhibition. As a rule, anti-HA2 MoAbs were poor haemolysis inhibitors.

Animals↗

[Expression of HLA antigens using the radioimmunoanalysis method].

Expression of HLA antigens class II on stimulated T lymphocytes was studied by radioimmunoassay. The products of all three HLA loci --DR, --DP, --DQ were present on stimulated cells in contrast to resting control T lymphocytes which do not express these antigens and are Ia negative.

HLA-D Antigens↗

Changes in the influenza virus haemagglutinin at acid pH detected by monoclonal antibodies to glycopolypeptides HA1 and HA2.

Monoclonal antibodies (Mabs) specific to the HA1 and HA2 subunits of the influenza virus haemagglutinin (HA) were used to show that changes in the antigenicity of the HA molecule at acid pH involve both HA subunits. In solid phase RIA (intact virus adsorbed) the acid-induced change was detected in the form of greatly increased binding of anti-HA 1 Mabs (IVA 1 and IVG 6) and anti-HA2 Mab (IIF 4). This increased binding could be most probably explained by alterations in accessibility of epitopes to the corresponding Mabs. Other Mabs examined (including 7 anti-HA2 Mabs specific to 3 independent antigenic sites) had either similar reactivities with both untreated and pH 5-treated virus or slightly but significantly increased binding to pH 5-treated virus. No effect of pH 5 treatment on antibody binding was observed with purified BHA in solid phase RIA. Nevertheless a similar pH 5-induced conformational change in the isolated BHA (like in intact viral HA in solid phase RIA) was detected in competitive binding assay carried out in liquid phase.

Antibodies, Monoclonal↗

Monoclonal antibodies to glycopolypeptides HA1 and HA2 of influenza virus haemagglutinin.

Anti-haemagglutinin monoclonal antibodies were prepared and their HA1 or HA2 specificity was determined by solid phase radioimmunoassay (RIA) using purified viral haemagglutinin (HA) and haemagglutinin glycopolypeptides HA1 and HA2, by radioimmunoprecipitation followed with SDS-PAGE, by immunoblotting and by inhibition of virus-induced haemagglutination. The capacity of these methods to estimate HA1 or HA2 specificity of anti-HA monoclonal antibodies (MoAb) was compared. HA1 specificity was demonstrated for all hybridomas originating from lymphocytes of mice immunized with complete influenza virus, except IIF4 hybridoma which was HA2-specific. All hybridomas obtained with lymphocytes from mice immunized with HA glycopolypeptide HA2 were HA2-specific. Anti-HA2 MoAb neither inhibit haemagglutination induced by the virus or by HA subunits nor neutralized viral infectivity, either alone or in mixture. As expected, all anti-HA1 MoAb were H3 subtype-specific, showing usually good reactivity only with viruses close to the virus strain used for immunization. Two anti-HA1 MoAb (IVA1 and IVG6) showed unusual cross-reactivity within the H3 subtype. All anti-HA2 MoAb were broadly cross-reactive within the H3 subtype. Moreover, a half of them showed high cross-reactivity with influenza viruses of the H7 HA subtype. But the same antibodies did not react with HA of H1, H2 and H8 subtypes.

Animals↗

Rescue of presumptive viral information from human cells by a helper oncovirus.

We have attempted to rescue presumptive human endogenous retrovirus(es) by using a competent animal oncovirus as a helper. Human melanoma cells (line HMB2) were fused, using polyethylene glycol, with mouse NIH-3T3 cells which had been infected and transformed by the Harvey murine leukaemia and sarcoma virus complex (MLV and MSV). The heteropolykaryons obtained were co-cultivated with fresh NIH-3T3 cells; filtered (Millipore 0.22 micron) medium from these was used to infect further NIH-3T3 cells. In these cells after several passages, vesicular stomatitis virus (VSV) pseudotypes could be produced. These were infectious not only for mouse cells (manifesting the helper MLV), but also for human cells (HeLa, HEC human embryo fibroblasts, HMB2); they were not infectious for CCL64 (mink) or for Vero (African green monkey) cells. The presence of such VSV pseudotypes infectious for human cells indicated that a human ecotropic virus [provisionally named rescued human virus (RHV)] had been rescued by the fusion of human melanoma cells with MLV-infected mouse cells. This was supported by the following evidence. The human-specific pseudotype was neutralized by sheep antisera raised to antigens selected by VSV from human tumour cell lines HMB2, T47D and HeLa. These antisera also aggregated NIH cells infected with MLV and RHV. Mouse antisera raised to antigens present in HIH cells infected with MLV and RHV, in contrast to sera raised to NIH cells infected with MLV only, immunoprecipitated an 85,000 mol. wt. protein band from human cells (HEC, HMB2 and HeLa) surface-labelled with 125I.

Animals↗

Characterization of influenza A-1983 epidemic strains by polyclonal and monoclonal antibodies and detection of two co-circulating antigenic variants.

Influenza virus strains isolated during 1985 epidemic in Czechoslovakia proved to be antigenically closely related to A/Bangkok/79, A/Philippines/2/83 and A/Texas/77 (all H3N2) viruses, if examined in haemagglutination inhibition (HI) tests with standard polyclonal antisera. If examined in HI tests with monoclonal antibody (MAb) IIB4, the virus isolates could be separated into two groups: those reacting to high titres (about two thirds of the isolates) and those negative with IIB4 (titre of less than 20; rest of the strains). A relationship to MAb IIB4 similar to that of freshly isolated A-H3 influenza virus strains was found with prototype strains A/Belgium/2/81 (highly positive with IIB4, HI titre up to 20 000 per 0.025 ml) and A/Philippines/2/82 (titre less than 20). Examination of the isolate labelled A/Prague/2/83, obtained from a single individual, suggested the existence of two stable and passage-independent lines of a single virus strain, namely one HI+ and the second HI- (highly positive and negative in HI tests with MAb IIB4, respectively). Solid-phase radio-immunoassay with 125I-labelled MAb IIB4 of the viruses under consideration showed that binding of virus with antibody had occurred in all cases and that, therefore, the negative results of HI tests with HI- strains were not due to the absence of binding of MAb IIB4 to the respective viral antigen.

Animals↗

A simple and rapid characterization of influenza virus isolates by monoclonal antibodies in radioimmunoassay.

Radioimmunoassay (RIA) with infectious allantoic fluid directly bound to solid phase, suitable for detection and further characterization of influenza virus isolates, is described. This simple and rapid method was applied for description of isolates obtained from different regions of Czechoslovakia during influenza epidemic in 1983. The results confirmed that all 13 examined isolates represent influenza A viruses possessing H3 subtype haemagglutinin very similar to haemagglutinin of influenza viruses A/Bangkok/1/79 (H3N2), A/Belgium/2/81 (H3N2) and A/Philippines/2/82 (H3N2), respectively.

Allantois↗

Analysis of influenza A virus neuraminidase using lectin test and monoclonal antibodies.

Influenza viruses causing epidemics in the U.S.S.R. in 1968-1982 and 1983 were analysed in the lectin test (LT) using polyclonal and monoclonal antibodies with specificity against neuraminidase (NA) of N2 subtype. Heterogeneity of the U.S.S.R. virus isolates in the reaction with monoclonal and polyclonal antibodies was demonstrated, though they were coming from the same year epidemic. The LT turned out to be an appropriate method to detect the antigenic drift in influenza virus NA. The results of LT were in a good agreement with those obtained by colorimetric estimation of the inhibition of enzyme activity and by competitive solid phase radioimmunoassay (SP RIA). In the LT only low steric inhibition of the NA with antihaemagglutinin monoclonal antibodies (MoAb) reacting in haemagglutination inhibition and virus neutralization tests was detected.

Animals↗

Indication and study of antigenic properties of influenza virus internal proteins by means of monoclonal antibodies.

Influenza A and B viruses isolated from animals and man were studied by solid phase radioimmunoassay (SP RIA) using monoclonal antibodies to the A/Dunedin/4/73 strain nucleoprotein (NP). Only influenza A viruses isolated before 1980 interacted with the monoclonal antibody set used, while strains isolated between 1980 and 1983 failed to do so. It was shown that RIA employing monoclonal antibodies was useful for rapid identification of influenza virus NP in the allantoic fluid containing fresh influenza A virus isolates.

Animals↗

Monoclonal antibody against an antigen selectively assembled into vesicular stomatitis virus virions from HeLa cells.

A mouse hybridoma cell line IIB9, secreting IgG2b antibody specific for a HeLa cell antigen, was obtained by fusion of a mouse myeloma cell line with spleen cells from mice immunized with purified VSV tsO45 mutant (defective in assembly of G protein) which had been reproduced at a non-permissive temperature in HeLa cells. The monoclonal antibody IIB9 was strictly specific for HeLa cells in two tests: (1) reaction with VSV or Chandipura virus phenotypically mixed with host cell antigen, (2) complement-dependent cytotoxicity test (51Cr-release).

Antibodies, Monoclonal↗

Purification of tick-borne encephalitis virus by affinity chromatography using monoclonal antibody.

The application of a simple technique for purification of tick-borne encephalitis (TBE) virus is described. TBE virus was grown in chick embryo cell (CE) cultures and the virus was concentrated by differential centrifugation. Final purification was made by the filtration through Sepharose column to which monoclonal antibodies to TBE virus had been bound. The method was effective in eliminating avian retroviruses.

Animals↗