Sidney Raffel

HEMOLYTIC ANTIBODIES FOR SHEEP AND OX ERYTHROCYTES IN INFECTIOUS MONONUCLEOSIS

Paul and Bunnell (1932) observed very high concentrations of agglutinins and hemolysins for sheep cells in the sera of patients during the acute stages of infectious mononucleosis, and this observation was found by these investigators to be of much value in the diagnosis of the disease. This finding has been confirmed and extended by Rosenthal and Wenkebach (1933), Boveri (1933), Bunnell (1933) and Bernstein (1934). In addition to the practical application in diagnosis, theoretical interest centers about the fact that antibodies for sheep cells are produced or enhanced in a disease of unknown etiology. Paul and Bunnell offered two explanations for their findings: (1) that the unknown agent responsible for infectious mononucleosis contains the heterophile or Forssman antigen; (2) that they were dealing with an example of isoagglutinin production elicited by abnormal cells, which are present either in the blood, or elsewhere, during active stages of the disease. In studies of infectious mononucleosis by different investigators, a probable bacterial origin has often been suggested, with special mention of the streptococcus group, diphtheroid bacilli, and Vincents organisms, all of which have been found in nose, mouth or throat lesions during the disease. Nyfeldt (1929) has reported the isolation of a small V-shaped or curved gram positive organism from the blood, which he called Bacterium monocytogenes hominis, and with which he produced the cellular blood picture of infectious mononucleosis in rabbits. Agglutination of this organism in 1 to 250 dilution by the serum of the patient was observed, but only five days after the temperature had become normal. Gorham, Smith and Hunt (1929) claimed to have produced the typical blood picture of the disease in guinea pigs by inoculation of membrane from the pharynx of

Complement-Fixation in Experimental and Human Poliomyelitis.∗

Conclusions The experiments presented above show that complement-fixing antibodies can be demonstrated in the blood of rats immunized with formalized Lansing virus, in convalescent monkey serum, and in some convalescent human sera when concentrated Lansing virus from cotton rats is used as antigen. The results with rat sera are complicated by the presence of antibodies against components of normal brain and spinal cord in both immune and normal animals, but the evidence indicates that the primary reaction with immune sera is between antibody and virus. Immunization as a result of injection of formalized virus is, therefore, associated with the production of serologically detectable antibodies to active virus. The experiments with the convalescent monkey and human sera provide definite proof that positive complement fixation can be demonstrated when concentrated Lansing virus prepared as described 2 is used as the antigen. These latter results further support the importance of the Lansing virus or an antigenically related strain as one responsible for the human disease.

Passive transfer of hypersensitivity to lepromin.

Conclusions These experiments indicate that the capacity to form a granulomatous response to lepromin can be induced in guinea pigs as a reaction of hypersensitivity directed toward M. leprae, and that this reactivity is transferable by viable lymphoid cells but not by heat-killed cells or by serum. On this basis, this responsiveness qualifies for inclusion in the category of the delayed hypersensitive states.

Sedimentation of Poliomyelitis Virus by Means of a Vacuum Ultracentrifuge.

This is a brief preliminary report of results indicating that we have been successful in sedimenting poliomyelitis virus, one of the smallest of the ultramicroscopic viruses, 1 from clear aqueous suspensions by means of ultracentrifugation in vacuum. The machine employed is similar in design to that recently described by Bauer and Pickels, 2 Thus far we have obtained complete results on 2 experiments. In both, the material subjected to ultracentrifugation consisted of a 25% suspension of glycerinated pooled virus cords in physiological saline. The lipoids in these suspensions were removed by ether extraction and the aqueous fraction was centrifuged in an Angle centrifuge at 3000 r.p.m. for at least an hour. In the first experiment the resultant fluid was water-clear to the eye; in the second, very slightly opalescent. Eight cc. of the clear suspension were placed in each of a series of celluloid tubes seated in the rotor. In the first experiment, the rotor was in motion for a total of 4 hours. Stroboscopic determinations indicated that for 2 hours of this period, the speed was between 27,500 and 30,000 r.p.m. After centrifugation, all of the tubes contained a small membranous type of sediment which could not be completely dispersed by repeated pipetting. The thicker central portion of the sediment presented a pinkish tinge apparently due to sedimentation of hemoglobin present in the original cord suspension. The supernatant was removed in 2 portions, the upper 6 cc. comprising the “top supernatant,” and the pooled bottom 1 cc. portions being the “bottom supernatant.” The sediment was resuspended in 6 cc. of saline. Monkeys were injected intracerebrally with 1 cc quantities of each of these fractions in varying dilutions. The results.

Pathogenicity of the Tubercle Bacillus.

Conclusions The Boivin type of antigenic complexes derived from the human strain (H37Rv) of M. tuberculosis appear to abet generalized infection in guinea pigs receiving very small numbers of organisms. It is suggested that such complexes may function as a “virulence factor” for the tubercle bacillus in a susceptible species of host. The possible relationship of such a factor to acquired resistance to the tubercle bacillus is under investigation.

THE SPECIFICITY OF PATHOGENICITY

This chapter discusses some findings on pathogenicity. General information concerning the potentially pathogenic species of enteric bacteria suggests that those virulent for man as judged by their isolation from the blood stream or from burned areas, are usually resistant to the bactericidal action of “normal” human serum, and also resist killing by the sera of unimmunized laboratory animals. However, a relationship between serum resistance and relative pathogenicity has been revealed partially by direct experiments. The question of relationship of serum resistance and sensitivity to high or low virulence is predicated upon two considerations. Resistance to serum might occur because of a structural feature of the cell wall, which makes the bacterium impervious to the lytic action of antibody and complement. Or, the resistance might be only apparent, depending upon the absence of appropriate “naturally occurring” antibodies in the serum of the subject tested.