Stanley Marcus

Detection by immunofluorescenge of antibodies specific for human malignant melanoma cells

Cells were separated from tissue specimens of human malignant melanoma and grown in tissue culture on coverslips. After 8 days of growth the explanted cells were fixed and stored until used. Sera from 15 patients with malignant melanoma were reacted with the fixed melanoma cells and a variety of other cells. The indirect immunofluorescence method was used to test for presence of antibodies. Sera from all 15 melanoma patients reacted with fixed melanoma cells giving intense fluorescence of intracytoplasmic granules. The fluorescent material appeared to be independent of melanin granules. Homogenate of fresh melanoma but not of other neoplastic or normal tissue absorbed out the anti‐melanoma antibodies. The human antimalignant melanoma antibodies did not react with live malignant melanoma cells or with fixed or live cells from benign nevi or fixed guinea pig kidney, human kidney or HeLa cells. The evidence suggests that these antibodies are unable to penetrate living cell membranes.

Latex agglutination reactions between human chorionic gonadotropin and rabbit antibody.

Summary It has been shown that rabbit antibody may be formed to the antigen, Human Chorionic Gonadotropin (Antuitrin S). Polystyrene latex particles were found to serve as an agglutinable carrier for this antigen-antibody system. The possibility of using this agglutination test for HCG as a suitable test for pregnancy is being explored.

Immunization of Mice with polysaccharides of Histoplasma capsulatum.

Summary H. capsulatum strain G17 (M) polysaccharide was capable of producing active resistance against intravenously induced histoplasmosis in mice. However, mice immunized with a H. capsulatum whole yeast cell antigen showed greater resistance when the H. capsulatum polysaccharide immunized, whole cell immunized and nonimmunized control mice were challenged by the intravenous injection of known numbers of viable H. capsulatum yeast phase organism.

Sporulation characteristics of Histoplasma capsulatum

Histoplasma capsulatum produces two types of aleuriospores (7), macroconidia and microconidia. The macroconidia are typically large tuberculate, spherical to pyriform spores described as ranging from 10--25# (13) or 8--14/ , (5) in diameter. These macroconidia are formed in the aerial portions of the mycelium generally at the end of short pedicels but may be sessile or formed at the elld of long hyphae (13). As described by HOWELL (7) the large aerial spores begin their development as bulbous enlargements on the ends of lateral branches. These branches may be simple with a single spore on the end of each branch, or one to several spores may be produced acropetalously and directly on a short branch. As these spore initials or prespores increase in size, they are cut off from the rest of the hyphae by a cross wall near the base, become spherical to pyriform in shape, and the walls gradually increase in thickness. The character of the spore wall varies greatly with the strain examined, the medium used or the part of the mycelium in which it is produced (7). In addition to tuberculate and smooth maeroconidia, there are forms having only severallarge bulbous or convoluted swellings or nmnerous spiney or warty projections (7, 11). Preponderantly smooth-walled macroconidia have beei1 noted deep in the mycelial mat next to or embedded ill the agar (7,11). Certain spores formed in these regions present a halo of substance SUl~ounding the body of the macroconidium and have been called nymbospores (12). The microconida are smooth-walled spores, spherical, pyriform

Macrophage and Lymphocyte Contributions in Resistance to Candida Albicans Infections

Model in vivo and in vitro experimental systems have been used to study the efficacy of specific and nonspecific immunization against Candida albicans infection induced in mice. Experiments were designed to compare the extent of resistance in specificity immunized, endotoxin treated and saline treated animals. In vitro phagocytic and postphagocytic killing (cytopepsis) of macrophages or lymphocyte-macrophage combinations from such animals were determined. In the in vitro experiments the macrophage systems destroyed the yeast cells more rapidly than did the lymphocyte-macrophage combinations. Since equivalent numbers of yeast cells were phagocytized, the differences observed were a function of cytopepsis of the organisms.

Effect of Influenza Virus Infection on Phagocytic and Cytopeptic Capacities of Guinea Pig Macrophages

Guinea pigs were exposed to influenza Al virus by aerosol. Peritoneal and alveolar macrophages were harvested three days after virus exposure and allowed to attach overnight to Leighton tubes. These macrophages were then challenged with a suspension of Klebsiella pneumoniae for two hours. The macrophages were washed free of extracellular bacteria and antibiotics were added to prevent extracellular multiplication. Plate counts were made at various time intervals on disrupted macrophages to determine the number of viable intracellular bacteria remaining. Alveolar macrophages that had been exposed to virus in vivo ingested the bacteria at a rate significantly greater (p <.05) than that of non-virus exposed control macrophages. However, virus exposed macrophages exhibited significantly reduced intracellular killing (cytopepsis) (p <.01) as compared to controls. In vitro virus exposed macrophages exhibited no significant difference in the rate of phagocytosis or cytopepsis. The data support the hypothesis that...

EFFECT OF PYROGEN ON PHAGOCYTIC DIGESTION AND SURVIVAL OF X-IRRADIATED MICE

Piromen in doses of 1 mu g per injection administered twice daily for 3, 5, or 7 days significantly increased intracellular digestion of chicken erythrocytes by mouse phagocytes. Similar treatment with 0.1 mu g of Piromen for 5 to 7 days also increased intracellular digestion. However, Piromen in doses of 0.1, 1, or 2 mu g per injection administered twice daily for various intervals prior to and after x irradiation failed to increase survival of x- irradiated mice. (auth)

ANTIGENS OF BLASTOMYCES

The material to be reported stems from interest in the hypothesis that reagents more specific than those available might be prepared for use in serologic and skin test reactions involving systemic mycotic pathogens. The problem of specificity of skin test reagents is of prime importance because of evidence demonstrating that cross reactions may occur among individuals sensitive to histoplasmin, blastomycin, coccidioidin, and Emmonsin.l-K* Martin has presented the results of wide-ranging studies on various Blastomyces dermatitidis antigens.6-8 Soluble antigen, obtained by sonic vibration of yeast-phase cells of B. dermatitidis, yielded as satisfactory a complementfixation test antigen as entire yeast-phase cells, but had the advantage of solubility and ease of storage. A polysaccharide precipitated from yeast-phase cell extract was found to be serologically active as well as capable of stimulating a limited degree of resistance to disease in mice. The crude extracts, however, were not found to be of significant value as skin test antigens. We have further explored these areas and, in this report, present aspects of strain variation in B. dermatitidis and the comparison of serologic, skin-reactive, and immunogenic properties of polysaccharides prepared from different strains of this organism.

Effects of Cortisone, Ascorbic Acid and Piromen on Phagocytosis in Mice.∗†

Summary Cortisone, ascorbic acid and Piromen were tested in mice for their effect upon macrophage activity as determined from the splenic uptake of colloidal ThO2 and upon the phagocytic activity of leukocytes in the peritoneal cavity against Micrococcus aureus. Cortisone in high doses (0.1 to 1 mg every 12 hours) significantly enhanced phagocytic activity of both macrophages and leukocytes. Ascorbic acid in doses as high as 1 mg every 12 hours had no effect on the activity of macrophages but this dose did significantly enhance the phagocytic ability of leukocytes in the peritoneal cavity. Piromen (0.1 μg every 12 hours) significantly increased the phagocytic activity of both types of cells. However, doses above or below this value were without significant effect in either type of experiment. The more potent auxophagocytic agents (cortisone and Piromen) were without salutary effect when tested singly and in combination in protection experiments in mice challenged with predetermined doses of Klebsiella pneumoniae.

Effect of properdin on whole body irradiated mice and rats.

Summary Under conditions employed postirradiation treatment of mice and rats with partially purified bovine properdin and purified human properdin administered intraperitoneally or intravenously did not afford protection against the effects of whole body x-irradiation. There appeared to be a relationship between delayed mortality and time of injection.