An attempt to determine the relation between levels of IgG, IgA and IgM immunoglobulins and titers of antibodies against RS virus.

In a group of 126 children aged 5-24 months with respiratory infections, the relation between levels of IgA, IgG and IgM immunoglobulins and titers of antibodies against the RS virus were studied. In children aged 5-12 months with high titers of antivirus antibodies, elevated levels of IgG were more frequent, but high antibody titers were not related to IgA and IgM immunoglobulins.

Effect of poly I:C on cell proliferation in vivo.

Female Swiss albino mice were injected intraperitoneally with poly I:C1 microgram/g body weight. The animals were killed 2, 12, and 24 hr after the injection of poly I:C. Half an hour before the death of animals they were reinjected intraperitoneally with 3H-thymidine. Fragments of ileum were excised. The quantification of autoradiographs included the counting of labeled cells, mitotic cells as well as the recording of cell position along the ileal crypts. The spatial distribution of both the mitotic and DNA-synthesizing cells of colonic crypts were affected by poly I:C at neither of time intervals between the injection and the death of animals. On the other hand, the parameters of cellular proliferation, i.e. mitotic and labeling indices, decreased significantly 12 and 24 hr after the injection of poly I:C, being unaffected 2 hr after the injection.

Effect of interferon and poly I: C on DNA fiber propagation in L 929 cells.

Autoradiographic analysis of DNA fiber replication was used to measure the rate of DNA propagation, the distances between initiation points of adjacent replicons and the direction of DNA propagation in L929 cells treated with either interferon or poly I: C, or untreated. Poly I:C ceased by 44% the rate of DNA propagation while the interferon had no effect. Poly I:C but not interferon reduced the mean distance between initiation points by 32%.

Influence of theophylline on interferon production and cAMP level Lpa cells.

In the presence and absence of methionine initiator tRNA (tRNA(iMet)) theophylline applied for 1 h starting 6 h after induction increased significantly (2-fold) interferon (IFN) yield. Theophylline did not change the kinetics of IFN production in the presence of tRNA(iMet). The shut-off stage was present and the production was prolonged to 24 h after IFN induction. In comparison with the values obtained in untreated cells, poly I:C plus DEAE dextran (inducing complex) caused an about 2-2.5 fold increase of cyclic adenosine 3'5'-monophosphate (cAMP) level. The similar results were observed in the cells treated with inducing complex plus tRNA(iMet). About 2-fold higher cAMP level occurred in the cells under the influence of the inducing complex plus theophylline in comparison to the level obtained without theophylline. The addition of tRNA(iMet) did not influence the above presented data.

Stability of the lyophilized F(ab')2 fragments of horse tetanus antibodies isolated by affinity chromatography.

F(ab')2 fragments of horse tetanus antibodies were obtained from horse hyperimmune sera after peptic digestion. The digest was passed through a column of tetanus toxoid coupled with Sepharose 4B, F(ab')2 fragments were eluted with a solution of 5 mM HCl in 150 mM NaCl and the eluates were concentrated by ultrafiltration and lyophilized. Glycine and human serum albumin were used as stabilizing agents. Polyacrylamide gel electrophoretic mobility and molecular weight of the fragments remained unchanged after lyophilization. Freeze-dried preparations stored two months at 56 degrees C showed only a slight decrease in antitetanus activity. The ORD measurements and spectrophotometric determinations of unfolding over pH range 2.1-5.0 show that the F(ab')2 fragment structure is highly stable in acid medium.

A study of human embryonic cells treated with UV-inactivated measles virus.

Some features of persistent infection were observed in human embryonic fibroblast cells (CLV21K cells) after single treatment with UV-inactivated measles virus. Morphological changes and chromosomal abnormalities were found. In subsequent passages of CLV21K cells, viral antigens were discovered by rosette haemadsorption, immunofluorescence tests and electron microscope or scanning electron microscope studies. No infectious virus was found.

Anti-pseudomonas immunoglobulin. II. Preparation of purified sheep immunoglobulins.

Several methods of fractionation of sheep anti-sera against Pseudomonas aeruginosa were applied for the preparation of immunoglobulins. The methods involving salting out with ammonium sulfate or purification by means of caprylic acid were shown to be most suitable for isolation of immunoglobulins under manufacturing conditions. The preparations obtained by these methods agglutinated Pseudomonas aeruginosa, suspension in vitro and protected mice against lethal infection with this microorganism.

Laboratory-scale procedure for preparation of horse anti-dog lymphocyte globulin (HADLG).

Procedure of preparation of horse anti-dog lymphocyte globulin on laboratory scale is described. Adsorption of antibodies against dog plasma proteins was performed with the use of immunosorbent prepared by coupling dog plasma proteins to aluminium hydroxide gel. Anti-lymphocyte immunoglobulins G and T were purified using two-step fractionation. Addition of glycine to the purified immunoglobulins prevented the aggregate formation. Purified immunoglobulins were tested in dogs receiving kidney allografts. The short term administration of HADLG significantly prolonged the time of survival of allografted kidney in comparison to the untreated controls.

The influence of tRNA on interferon induction by poly I:C in Lpa cells.

The kinetics of interferon production, induced by poly I:C in the presence of DEAE dextran, was studied in Lpa cells. Optimal interferon production was obtained at concentration of 10 micrograms/ml poly I:C and 200 micrograms/ml DEAE dextran. Transfer RNA applied to Lpa cells during induction and synthesis of interferon resulted in continuous production of interferon for 24 hrs. The application of tRNA to cells during the shutoff of interferon synthesis had no effect on the kinetics of interferon production.

Anti-Pseudomonas immunoglobulin I. Immunization of sheep.

The production of sheep Pseudomonas immune serum is described. The animals were immunized with Pseudomonas vaccine prepared from strains of Pseudomonas aeruginosa belonging to seven immunotypes of Fisher's schema. Immune sera obtained from hyperimmunized sheep are characterized by a high level of stabel antibodies with a good protective activity for mice against infection with Pseudomonas aeruginosa. It has been demonstrated that sheep may be used for a long time as a source of immune serum.

Preparation of rubella virus haemagglutinating antigen by polyethlene glycol precipitation.

Polyethylene glycol (PEG) was used for precipitation of rubella virus haemagglutinating (HA) antigen from the fluid phase of infected Vero cell cultures, maintained with or without calf serum. In both cases potent antigen preparation were obtained. When the antigen was precipitated from serum-containing medium, higher titres were obtained, provided that substrate or final product were treated with Tween 80 and ether. HA antigen obtained by PET precipitation was entirely valuable in the haemagglutination-inhibition test, as compared with antigens prepared by other methods.

Inhibition of vaccinia virus replication in RK-13 cells by N,N'-bis(methylisatin-beta-thiosemicarbazone)-2-methylpiperazine.

N,N'-bis(methylisatin-beta-thiosemicarbazone)-2-methylpiperazine in a 100 mumol/l concentration inhibited the reproduction of vaccinia virus in RK-13 cells by about 90%. This compound (bis-IBTMP) had no influence on virus adsorption and on early stages of virus multiplication, but affected virus reproduction from 12 to 24 hr post-infection (p.i.). The incorporation of 3H-thymidine into infected cells increased during first 10 hr p.i., decreasing gradually afterwards. In the infected cells treated with bis-IBTMP the same tendency was observed up to 10 hr p.i., but later on the incorporation level remained unchanged. The uptake of 14C-amino acids in the presence of bis-IBTMP was reduced both in vaccinia virus-infected and non-infected RK-13 cells.

Purification of vaccinia virus by zonal centrifugation and analysis of viral protein composition.

Vaccinia virus propagated in rotated cultures of RK13 cells was purified by sucrose density gradient zonal centrifugation. The purified virus was dissociated with sodium dodecyl sulfate end its protein composition analyzed by polyacrylamide gel electrophoresis. The protein composition of the virion envelope, isolated by the non-ionic detergent Nonidet P-40, was also determined.

Effect of streptovirudin on parainfluenza 3 virus multiplication.

Streptovirudin (SV) in a concentration of 100 microng/ml reduced the multiplication of parainfluenza 3 virus by about 3.5 log units. When uninfected WISH cells were treated for 2 and 12 hours with 100 mug/ml of SV, 3H-thymidineincorporation was decreased, but no marked differences were observed in the incorporation of 3H-uridine. The level of intracellular cyclic adenosine monophosphate (c-AMP) was found to decrease in SV-treated cells.

Effects of plant transfer ribonucleic acids on interferon production.

An optimal interferon (IFN) production was obtained at concentrations of 50 micrograms/ml poly I:C and 2000 micrograms/ml DEAE dextran in Lpa cells. It was shown that methionine initiator tRNA (tRNAiMet) in a dose 50 micrograms/ml or crude tRNA (tRNAc) applied to Lpa cell during the stages of IFN induction, IFN induction and synthesis, as well as during IFN synthesis resulted in a continuous IFN production for up to 24 hr. Exposure of the cells to 150 micrograms/ml tRNAiMet during the stages of IFN induction and IFN induction and synthesis caused total inhibition of IFN production. This effect was partially observed only after the high dose of tRNAc. Addition of a high dose of tRNAiMet or tRNAc to cells during the synthesis stage caused no inhibition but prolongation of IFN production.

[Dipyridamole as an inhibitor of vaccinia virus replication]. / Dipiridamol kak ingibitor razmnozheniia virusa vaktsiny.

Dipyridamole in concentration of 25 microM inhibited the multiplication of vaccinia virus in about 90% of cells. In the presence of this substance, [3H]-uptake was sharply reduced both in uninfected and infected RK13 cells, while [14C]-uptake was not inhibited incorporation of [3H]-thymidine and [14C]-amino acids into viral particles. The present findings suggest that the antiviral character of dipyridamole is related with the inhibition of the substrate transport through the cell membrane.

[Polish preparations for preventing and treating infections caused by Pseudomonas aeruginosa]. / Pol'skie preparaty dlia profilaktiki i lecheniia infektsii, vyzvaemykh Pseudomonas aeruginosa.

Three preparations, viz. sheep serum for local application ("Sanoserum"), anti-Pseudomonas sheep immunoglobulin for passive immunization ("Immunoglobulin Pseudomonas") and Pseudomonas vaccine for active immunization ("Pseudovac"), have been obtained at the Central Laboratory of Sera and Vaccines in Warsaw, Poland. The laboratory study and clinical trial of these preparations have been carried out. The preparations have proved to be effective as supplement to the conventional methods for the early diagnosis and treatment of P. aeruginosa infection in burn patients.