The Journal of Experimental Medicine

Coburn, Alvin F.; Graham, Claire E.; Haninger, Joan

doi: 10.1084/jem.100.5.425pmid: 13211905

Whole egg yolk incorporated as a supplement in the diet of baby guinea pigs afforded protection against anaphylactic arthritis as determined: ( a ) by measurement of joint swelling; ( b ) by the rise in serum level of some substance which reacts with diphenylamine; ( c ) by histologic examination. The active material in egg yolk was shown to be in the alcohol-soluble fraction. Attempts to identify the active material with any known lipid have to date been unsuccessful. In the screening of lipid substances for protection against anaphylactic arthritis, it is shown that the weanling guinea pig is suitable, that a 3 week period is adequate, and that the test substance can be administered satisfactorily in the diet. Footnotes Submitted: 15 May 1954

Ackermann, W. Wilbur; Rabson, Alan; Kurtz, Hilda

doi: 10.1084/jem.100.5.437pmid: 13211906

A detailed study of the cytological changes which are induced in HeLa cells by the Saukett strain of Type III poliomyelitis virus has been made. The observations were of cultures in which a single sequence of infection was induced. The cytological changes were examined in relation to the growth curve of the virus in the same type of culture. This curve showed a latent period of 4 to 5 hours, followed by a gradual release of virus over an interval of 6 to 7 hours. Changes in the staining character of the cells occurred before the major portion of the viral yield appeared. The infected cells exhibited a striking cytopathology with increased basophilia, nuclear pyknosis, and basophilic cytoplasmic granules. Individual cells showed characteristic differences in the rate at which the cytopathology progresses. The multiplication of the virus in HeLa cells was inhibited by fluorophenylalanine. The inhibitory effectiveness of the antimetabolite was related to the age of the infection. It apparently inhibits only an early stage of viral development. The inhibition is completely reversed by phenylalanine if the amino acid is added within 6 hours, not later, after the induction of virostasis. The data are interpreted in terms of the rate at which the ability of the infected cell to support viral synthesis was lost. Flurophenylalanine also inhibited the multiplication of HeLa cells; however, the effect upon the uninfected cell was reversible after 3 days, as indicated by viability after such treatment. While the fluoro derivative completely inhibited viral multiplication, it did not prevent the cytopathogenic effect of the virus. In the presence of fluorophenylalanine, the disintegration of an infected cell proceeded at what appeared to be the ordinary rate, without any increase of the infectious agent. Experimentally the processes leading to viral increase and to cellular injury have been shown to possess a significant degree of autonomy. Footnotes Submitted: 25 June 1954

Waksman, Byron H.; Porter, Huntington; Lees, Marjorie D.; Adams, Raymond D.; Folch, Jordi

doi: 10.1084/jem.100.5.451pmid: 13211907

Fractions of bovine white matter, prepared by the methods of Folch and Lees, were studied for chemical composition and for their ability to produce experimental allergic encephalomyelitis in rabbits. Evaluation of the disease and of the lesions in animals injected with the more active fractions at several dose levels permitted comparison of the antigenic activity of these materials. When tissue was fractionated by the methods of Folch and Lees, antigenic activity was found in the chloroform-methanol extract but not in the denatured tissue residue. This activity was traced to proteolipides A and B and to the lower phase, more specifically the ether-soluble fraction of the lower phase. Proteolipide C was inactive. Correlation of the chemistry of fractions with their antigenic activity suggested two possibilities: ( a ) that there might be two antigens, one proteolipide and the other non-proteolipide; or ( b ) that a small specific proteolipide is responsible for all the observed activity. The high concentration of acetal phosphatide in the ether-soluble lower phase suggested that compounds of this type might be the hypothetical non-proteolipide antigen, but this hypothesis was disproved by analytic study of active and inactive materials. The possibility that proteolipide might account for all the antigenic activity was strongly supported by the experimental finding that total lipide and proteolipide progressively lost activity as proteolipide was degraded by adequate processing. The use of an entirely different method for preparing total lipides free of proteolipide (the colloidal iron technique) indicated that this loss of activity did not result from incidental removal of some non-proteolipide antigen. These tentative conclusions are in agreement with those of Tal and Olitsky and provide a satisfactory interpretation of the findings of Goldstein et al . The very fact, however, that the suggested proteolipide antigen would amount to no more than 1 per cent of the total chloroform-methanol extractives leaves open the possibility that some unrecognized trace substance may be the antigen. Skin tests with the various fractions indicated some cross-reactivity between proteolipides A and B and the ether-soluble lower phase fraction and a fair correlation of positive skin reactions with disease. This finding is compatible with the suggestion that the same antigen is present in both of these types of material. When the disease produced by whole tissue or fractions was evaluated by the use of the proportion of animals developing disease, the day of onset, and the severity of the histologic lesions, it was found that fractions produced milder disease of later onset than intact tissue at all dose levels. The disease-producing activity was not enhanced by increasing the dose; i.e ., it appeared to reach an asymptotic maximum below that obtainable with whole fresh tissue. This finding suggests both a quantitative loss of activity and a qualitative change during the initial chloroform-methanol extraction, a procedure which denatures all proteins in the tissue residue. A comparable change appeared to occur in whole white matter stored at –15°C. for 15 months and thawed and refrozen several times during this period. The later fractionation steps resulted in no apparent loss of antigenic activity. A scoring method employing the same type of data to estimate the actual relative antigen contents of different preparations is presented in the Appendix. Footnotes Submitted: 28 June 1954

Gray, Alan; Scott, T. F. McNair

doi: 10.1084/jem.100.5.473pmid: 13211908

The growth cycle of the virus of herpes simplex in chick embryo liver has been shown to follow the same pattern as in the chorioallantoic membrane and the rabbit's corneal cells. However, there is considerable variability in the time taken for the yolk sac-inoculated virus to get from the yolk sac into the liver. A brief description has been given of various fractionation procedures employed for obtaining isolated nuclei. It has been shown that free virus is not selectively adsorbed to isolated nuclei. Evidence has been presented to show that in the herpes-infected chick embryo liver, large proportions of the total virus can at times be found associated with the nuclear fraction. The percentage of the total virus in the nuclear fraction varies inversely with the titer of virus in the whole liver, and the number of hours after inoculation of the virus; only a negligible amount (as compared with that in the total) being associated with the nuclear fraction when a period of over 12 hours has elapsed after reappearance of virus. Furthermore, demonstration of virus in the isolated nuclei following extraction with hypertonic NaCl provides additional evidence that this virus is intimately associated with the nuclei. Footnotes Submitted: 29 June 1954

Kuhns, William J.

doi: 10.1084/jem.100.5.485pmid: 13211909

The method of filter paper electrophoresis was used to study proteins and protein-bound polysaccharides in sera obtained from subjects before and after a single booster dose of diphtheria toxoid, and in sera from allergic subjects. The electrophoretic patterns of precipitating antitoxic sera resembled those found in normal non-immune sera. However, skin-sensitizing antitoxic sera were distinguished by a relatively large beta globulin component and a small or indistinct alpha 2 globulin. Fusion of both components was present in some sera containing this variety of antitoxin. Considerable amounts of serum-bound polysaccharides in these sera migrated relatively slowly in contrast to the behavior of polysaccharides of precipitating antitoxic sera which migrated faster when tested under similar conditions. Alterations in proteins and carbohydrates were most readily observed in specimens containing high titers of antitoxin. There were no demonstrable differences between the electrophoretic behavior of sera obtained from subjects before or after immunization with toxoid. Electrophoretic patterns of serum from allergic subjects who developed marked eosinophilia showed attenuation of the alphas globulin associated with a relative preponderance of slow migrating protein-bound polysaccharides. These alterations were not present in sera obtained from the same persons before and after the development of eosinophilia. Changes in the proteins and polysaccharides could not be demonstrated with consistency in subjects with mild to moderate hay-fever symptoms. One person who developed severe acute hay-fever symptoms showed alterations in the beta and alpha 2 globulins. Rheumatic fever subjects showed no unusual changes in the distribution of serum components. However, transition from the acute process to convalescence is graphically demonstrated by the marked decreases in gamma and alpha globulins and in protein-bound carbohydrates. Footnotes Submitted: 20 July 1954

Maurer, Paul H.

doi: 10.1084/jem.100.5.497pmid: 13211910

It has been demonstrated that there are normally occurring antibodies to gelatin in human sera. Immunization with gelatin can in many cases increase the antibody level. The presence of these antibodies does not result in cutaneous reactions of the wheal and erythema type after injection of antigen. Many of the properties of the gelatin-antigelatin reaction and the precipitates formed are those of a truly specific antigen-antibody aggregate. Explanations have been advanced both as to the possible sources of the gelatin antibody and its significance. Footnotes Submitted: 30 June 1954

Maurer, Paul H.

doi: 10.1084/jem.100.5.515pmid: 13211911

Normally occurring antibodies to gelatin have been demonstrated in sera from many species. Immunization of rabbits with gelatin has been successful when adjuvant techniques were employed. Footnotes Submitted: 30 June 1954