The Journal of Experimental Medicine

Fong, Jacob; Chin, Dennis; Akiyama, Hajime J.; Elberg, Sanford S.

doi: 10.1084/jem.109.6.523pmid: 13654626

Studies of the relationship of protective serum factor to cellular resistance and to tuberculin skin sensitivity have demonstrated that protective serum factor may exist independently of a high level of cellular resistance, and that both protective factor and cellular resistance may be demonstrable without a concomitant hypersensitivity of the delayed type. The experiments with absorbed sera and the globulin fraction of immune serum indicated no specific association of protective serum factor with antibody globulin. The protective factor in immune serum was found to be thermostable and non-dialyzable. In vitro exposure of virulent tubercle bacilli to the immune serum from BCG-immunized animals failed to alter the bacterial capacity for destruction of monocytes and for intracellular proliferation. In vitro cultivation of normal and immune monocytes in normal or immune serum was not effective in changing the native susceptibility or resistance of these cells. Effective manifestation of resistance to virulent tubercle bacilli by immune monocytes was found to require the continuous presence of immune serum. The intracellular passage of virulent tubercle bacilli in an immune system (immune monocytes cultivated in immune serum) resulted in a decreased bacterial potential for destruction of normal monocytes when these were cultivated in the presence of immune serum. Footnotes Submitted: 28 December 1958

Liu, Chien; Eaton, Monroe D.; Heyl, James T.

doi: 10.1084/jem.109.6.545pmid: 13654627

By using the indirect method of fluorescent staining to study the antibody response in patients with primary atypical pneumonai associated with the development of cold agglutinin, it was found that the PAP antibody developed during the 2nd and 3rd week of the illness, and persisted for over a year, and is not related to the cold and streptococcus MG agglutinins. The development of the PAP fluorescent staining antibody paralleled the neutralizing antibody for the PAP virus as tested in cotton rats. The sensitivity of this specific serological test was indicated by the observation that 67 to 92 per cent of the patients in several outbreaks of PAP showed a rise of antibody titer during convalescence. Absorption of the sera with various tissue powders did not affect the PAP antibody detected by this method. Footnotes Submitted: 15 February 1959

Gross, Jerome

doi: 10.1084/jem.109.6.557pmid: 13654628

The skin of severely scorbutic guinea pigs which were losing weight contained no detectible neutral salt-extractible collagen. Conditions of growth (weight gain) which actively induced the formation of neutral salt-extractible collagen in the skin of normal guinea pigs failed to do so in the animal with ascorbic acid deficiency. No excess of non-collagenous proline was found in neutral salt extracts of scorbutic skin as compared with normal. Fractionation of these extracts failed to reveal the presence of significant amounts of a soluble component containing unusual proportions of glycine and proline relative to hydroxyproline. It is concluded that deficiency of ascorbic acid either interferes with the synthesis of new collagen in intact skin or causes its destruction and removal as rapidly as it is produced. Footnotes Submitted: 20 January 1959

Russell, P. S.; Gittes, R. F.

doi: 10.1084/jem.109.6.571pmid: 13654629

1. A reliable method of parathyroid grafting in rats, and of assessing the functional state of such grafts in terms of blood calcium has been devised. 2. Parathyroid homografts between rats of proven genetic diversity may be rejected as promptly as skin; but the majority survive for an indefinite period, there being no intermediate class. 3. Surviving parathyroid grafts in this genetically diverse combination invariably succumb in the presence of the immunity induced by a later skin graft from the same donor. 4. Parathyroid grafts between rats of less genetic diversity have usually responded to skin graft-induced immunity in the same manner as those between more dissimilar rats. The less divergent group, however, revealed an occasional persistent functioning of parathyroid grafts even in the face of the sensitivity incidental to an accelerated reaction to skin. 5. The hypothesis is presented that the parathyroid tissue of the rats studied was deficient in effective transplantation antigens as compared with those of their skin. This weakness in apparent antigenicity may be a reflection of a deficiency in production, availability, or content of antigens in the tissue transferred. Footnotes Submitted: 1 January 1959

Slade, Hutton D.; Kimura, Yoshitami

doi: 10.1084/jem.109.6.589pmid: 13654630

Heat-killed cells of Group A streptococci caused death of the adrenalectomized rat. While the adrenalectomized rat readily succumbed to intraperitoneal infection with living cells, death was due primarily to toxicity. The normal rat was highly resistant under either condition. For studies on the toxic materials, the cells of numerous serological types of group A streptococci, and of a Group B and a Group D streptococcus, were extracted with 0.1 N HCl at 100°C. or by sonic oscillation. The extracts, containing macromolecular components, were subjected to chemical fractionation and purification. C substance and M protein of Group A streptococci released from the cell by sonic oscillation were toxic to the adrenalectomized rat in quantities of 1 mg./100 gm. rat. Death usually occurred within 2 hours. On the other hand, C substance and M protein released from the cell with HCl at 100°C. were relatively non-toxic to the adrenalectomized rat. The sonic-extracted C substance of streptococcal Groups B, C, and D was also toxic. The toxic property of the C and M preparations was neutralized in vitro in each case by group and type-specific rabbit antiserum. Heterologous antiserum was without effect. Adrenalectomized rats which received homologous antiserum 18 hours before challenge were also resistant to the toxicity of the C and M preparations. Trypsin destroyed the toxic effect of the M protein preparations and was without effect on the toxicity of the C substance. The R antigen and a nucleoprotein component of Group A streptococci, preparations of protein from Groups B and D streptococci, and coagulase from Staphylococcus aureus were all found to be essentially non-toxicic for the adrenalectomized rat. Large quantities of peptone, crystalline albumin, and rabbit serum were also without effect. Footnotes Submitted: 4 March 1959

Keeler, Richard F.; Lovelace, Stuart A.

doi: 10.1084/jem.109.6.601pmid: 13654631

The urinary excretion of silicon in the rat was found to be enhanced beyond normal levels by the administration of various chemical forms of silicon. The excretion was enhanced to a much greater degree by the administration of ethyl silicate than by magnesium trisilicate, sodium metasilicate, or water glass. The tolerance level of rats to sustained daily doses of ethyl silicate fed via stomach tube was approximately 15 to 30 mg. of silicon per rat per day. Urinary silicon excretion was found to be a straight line function of the concentration of ethyl silicate administered, via stomach tube, with approximately 18 per cent of the administered silicon appearing in the urine at all levels tested. Using sustained dietary additions of ethyl silicate as a means of enhancing urine silicon levels, artificial siliceous urinary calculi were consistently produced on zinc pellets implanted in the bladders of rats. Footnotes Submitted: 16 January 1959

Fedoroff, S.; Cook, B.

doi: 10.1084/jem.109.6.615pmid: 13654632

When NCTC clone 929 (strain L) cells were grown either continuously or at intervals in toxic human serum, they became resistant to the toxic substance or substances. The resistance developed to toxic human serum was passed from one cell generation to another; i.e ., it was heritable. The resistance to toxic human serum, developed in strain L cells, was associated with a decrease in their ability to adsorb toxic substances from the serum. This suggests that there is a decrease in the number of loci in the resistant cells where toxic substances can be bound. The resistant cells differed from the sensitive cells in their morphological appearance, the increased frequency of minute chromosomes, and the increased adhesion to glass surfaces. The cells which developed a resistance to a particular toxic serum also resisted the toxic effects of some other sera, but not of all. Footnotes Submitted: 25 January 1959

Rothbard, Sidney; Watson, Robert F.

doi: 10.1084/jem.109.6.633pmid: 13654633

Renal glomerular lesions were induced by rabbit serum containing antibody to rat collagen injected intravenously into rats prepared with subcutaneously administered Freund adjuvant. Neither the anti-collagen serum nor the adjuvant alone induced the lesion. The lesions were characterized by diffuse glomerular injury with swelling, shredding, and fusion of the basement membranes, crescent formation, cellular proliferation, numerous multinuclear giant cells, and capillary hyaline thrombi. Various rabbit antisera, including those against fish collagen or rat serum failed to induce the renal lesion when substituted for anti-rat collagen serum. Also, anti-rat collagen serum absorbed with its homologous antigen, native rat collagen, failed to induce the lesion. Although complete adjuvant, i.e . with mycobacteria, in which normal serum was incorporated enhanced the glomerular lesion which resulted from intravenous injection of anti-collagen serum, the incomplete adjuvant without serum was sufficient. Comparison of the renal lesions induced by anti-collagen serum with nephrotoxic nephritis induced in rats by rabbit anti-kidney serum showed that they differ histologically. Also the antisera used to produce these two renal lesions differ immunologically. Antibodies to normal rabbit serum developed in rats injected intravenously with rabbit anti-rat collagen serum after preparation with adjuvant, but not when adjuvant was omitted. The pathogenesis of the renal injury is discussed as a manifestation of an antigen-antibody reaction, with nephritis occurring only after the adjuvant-stimulated antibody to the rabbit globulin has been formed in the rat and has reacted with the rabbit anti-rat collagen already fixed by its homologous antigen in the kidney. Footnotes Submitted: 15 February 1959

Fisher, Harold W.; Puck, Theodore T.; Sato, Gordon

doi: 10.1084/jem.109.6.649pmid: 13654634

Two purified serum protein fractions, fetuin and serum albumin, will replace whole or dialyzed serum in supporting the growth of single S3 HeLa cells in an otherwise chemically defined nutrient solution. In the serum-free medium, single S3 cells will form macroscopic colonies with essentially 100 per cent efficiency. The generation time of S3 cells in the serum-free medium is approximately 50 per cent greater than that observed in an optimal, serum-containing medium. All components of the serum-free medium are available commercially, except fetuin, which can easily be prepared in substantial quantities. The problem of the purity of the protein preparations and of their possible roles in promoting cell growth is discussed. Footnotes Submitted: 8 February 1959