Alvar A. Werder

Effect of skin extracts on the viability of homologous skin grafts in mice.

The purpose of these experiments is an attempt to prolong the survival of homografted skin on heterozygous CFW mice which had received subcutaneous injections of homologous skin extracts. Kohde,’ Allen,2 and Billingham, and Medawar? have reported on skin extracts prepared from homologous skin in animals of various species in an attempt to prolong the life of transplanted homografts. The results are reported with divergent success. Rhodes’sl work on the survival of homografts influenced by skin extract injections is not described in sufficient detail to warrant critical analysis. Allen and associates* have reported that the survival time of homografts in rabbits was extended 16 days by “desensitization” of the recipient with specific donor antigen. I t should be noted that these workers used phenol as a preservative in the preparation of the skin extracts. Billingham and MedawaP reinvestigated the above report and showed that “the lifetime of skin homografts transplanted to rabbits may be prolonged by the injection of a weak solution of phenol in saline. This prolongation of life is probably due to a nonspecific reaction to stress.” These workers were unable to prolong the survival of skin homografts by the injection of phenol-free donor skin extracts into the recipient rabbi IS.

Erythropoietic Stimulating Factor (ESF) as a Stimulant of Cell Growth in vitro

Summary Cultures of 2 cell lines have been tested for their response to administration of erythropoietin (ESF). The ability of ESF to stimulate growth of cultures of human synovial membrane cells and human monocytic leukemia cells was demonstrated. ESF is ineffective in stimulating cell culture multiplication in very rapidly growing cultures. Suggestion was made that ESF be called “Nonspecific Growth Factor” (NGF).

Electron Microscopic Quantitation of Viral Particles in Tissues of Leukemic Mice.

Summary A simple method for estimating the relative number of viral particles in thin sections of tissue is described. By this method particle frequency was defined as the presence or absence of particles within a specified sectioned area, which was dependent upon the number of nuclear sections examined with the electron microscope. This technique was applied to CFWW axenic mice inoculated with human leukemic cells. It was observed that the number of viral particles appeared to increase as the cells were serially transferred in mice.

Observations on human colonic cellular suspensions and filtrates as etiologic agents in ulcerative colitis

SummaryA preliminary experiment was designed to test resected colons from patients with ulcerative colitis for the presence of a possible causative agent.1. A cellular suspension and a cell-free filtrate prepared from diseased colon did not produce ulcerative colitis but were shown to be antigenic to rabbits by means of the Ouchterlony agar-diffusion technic. An antigen common to all colon filtrates was demonstrated. In addition, a second antigen, common only to diseased colon filtrates and not present in normal colon filtrates, was observed.2. The cell-free filtrate from diseased colons produced an unidentified precipitate in human amnion-cell culture, whereas normal colon filtrates exhibited no visible effect; neither resulted in a cytopathogenic effect on the cell cultures.3. Formation of this unidentified precipitate in cell culture could be prevented by adding specific rabbit antiserum to the filtrates prior to inoculation of the cell cultures.

Distribution of Virus-Like Particles in the Lymphatic Tissues of “Nonleukemic” CFWW Conventional Mice.

Summary A study was made on the “non-leukemic” conventional CFWW mice which show a 29% incidence of spontaneous leukemia by 24 months of age. An estimate on the presence of virus particles in the thymus showed 61% with C type particles and 28% with A 2 particles. In addition, 22% of the spleens and 17% of the lymph nodes examined contained C type particles. The C type particles in the thymus were associated with epthelial cells while the A 2 particles were within lymphocytes. This particle concentration within certain cell types, as it may relate to sampling error, was discussed. The implication of the thymus as a source of virus was discussed, including the possible role of the thymic blood barrier and epithelial cells in the “release” of particles to other lymphatic tissues.

Tissue culture of the sera in human ulcerative colitis.

1. Control human sera had no effect on human amnion cell culture. 2. The sera from nineteen of twenty-one patients with ulcerative colitis had a marked cytopathic effect on cell cultures, suggesting an autoimmune effect. The sera of three of these showed an additional cytopathic effect on human amnion cell culture, suggesting a viral-like quality. 3. The sera from patients after total colectomy and protectomy for ulcerative colitis had no effect on cell cultures. 4. Rabbit antisera did not produce demonstrable antibodies or protect against the cytopathic effect in cell culture.

Erythropoietic stimulating factor (ESF) associated with an oncogenic virus-host system in vitro.

Summary The possibility that erythropoietic stimulating factor (ESF) might be involved in the relationship between tumor and virus was tested in vitro. Four experimental situations were studied: (A) Adenovirus, type 12 inoculated into hamster cell culture, (B) Adenovirus, type 4 into hamster cell culture, (C) Adenovirus, type 12 into monkey cell culture and (D) Adenovirus, type 4 into monkey cell culture. Each of these cultures was extracted and assayed for ESF. In addition, other cultures containing only virus or cells in medium or medium alone were extracted and assayed. Extracts of cultures of hamster cells inoculated with human adenovirus, type 12, were found to contain an ESF. This material was found only in small amounts in the other experimental situations. The occurrence of this ESF primarily in the experimental situation in vitro analogous to that in vivo which results in tumor formation suggests that an ESF-like material may be associated with the process of cell division in both situations.

Plaque Production with Infectious Bovine Rhinotracheitis (IBR) Virus.

Summary and conclusions Plaques were produced by IBR virus on bovine embryo kidney cells by both Dulbeccos agar-overlay and Postlethwaites liquid overlay methods. The lower plaque titers obtained when the latter method was used were perhaps due to errors in scoring plaques which were not clearly distinct. The formation of plaques by IBR virus was blocked by IBR hyperimmune serum in both the standard tube neutralization test and by incorporation of antiserum into agar-overlay. Plaquing of IBR virus on bovine embryo kidney cells provided a more quantitative means by which to study the host-range of this virus and a more precise system for the biological assay of infective IBR nucleic acids which may possibly be obtained from the virus.

Incorporation of P32 into Phosphatido-Peptide Fraction of Normal and Neoplastic Mouse Epidermis.

Summary Evidence was presented which demonstrates that the turnover of phosphorus in phosphatido-peptide is greater for neoplastic epidermis than normal epidermis. Histologically “normal” epidermis obtained from mice bearing a squamous cell carcinoma induced with 20-methylcholanthrene had a phosphorus turnover more closely resembling neoplastic than normal tissue. Further studies concerning the biochemical changes mentioned are in progress.

Anaphylactic Shock in Mouse II. Allergenicity of Water-in-Oil Emulsions With or Without Mycobacterium

Summary 1) Mice sensitized with 0.1 mg of bovine albumin incorporated into water-in-oil emulsion containing Mycobacterium butyricum showed anaphylactic reactivity when challenged 28 days postsensitization with as little as 0.01 mg of bovine albumin. 2) Mice sensitized in a similar manner exhibited signs of anaphylaxis when challenged 196 days following sensitization. 3) Anaphylactogenicity in mice of bovine albumin in emulsions lacking Mycobacterium butyricum was comparable to that in emulsions containing acid-fast bacilli.