The Journal of Experimental Medicine

Majno, Guido; Karnovsky, Manfred L.

doi: 10.1084/jem.107.4.475pmid: 13513913

Samples of normal grey matter, white matter, and peripheral nerves obtained from rats were incubated in Warburg vessels with glucose and a labelled lipide precursor (acetate, phosphate, choline, glycerol, glucose). The total lipides were then extracted and their radioactivity measured. The preparations were compared with respect to dry weight, lipide content, O 2 uptake, and ability to incorporate the various substrates into the lipides. Grey matter was found to be the least damaged by incubation, white matter the most. Damage to the tissue depressed lipogenesis to a greater extent than respiration. Five substrates were compared with respect to their degree of incorporation into the lipides of the various preparations. White matter, which had a greater oxygen uptake than peripheral nerves, showed the lowest degree of incorporation for most of the substrates studied. The results suggest that there are considerable quantitative differences in the metabolism of central and peripheral myelin. In the sciatic preparations, oxygen uptake and lipogenesis from acetate were found to decrease from the proximal to the distal end of the nerve. This finding may be relevant to the pathogenesis of peripheral neuropathies. The growth and metabolic activity of peripheral nerves were studied in rats aged 1 to 500 days, and the biochemical and histological findings were correlated. The results indicated that the lipogenetic activity of the Schwann cell was lowest in the newborn animal, and reached its peak at about 20 days. Comparative data were also obtained from the cerebral cortex. The growth pattern of peripheral nerves was distinctly different from that of the brain. With respect to changes in tissue weight, respiration, and lipogenesis, growing peripheral nerve correlated with body weight, while the brain matured much more rapidly. Footnotes Submitted: 6 August 1957

Garvey, Justine S.; Campbell, Dan H.

doi: 10.1084/jem.107.4.497pmid: 13513914

The retention of antigen in rabbit liver tissue, resulting from a primary intravenous injection, is influenced by immunization brought about by subsequent intravenous injections of the same antigen. In rabbits given a single primary intravenous injection of radioactive antigen, the retention of radioactivity in liver tissue, after a period of 21 days, was greater than when the primary injection was followed by secondary injections of the same, but non-radioactive antigen. The results were similar for both S 35 -azohemocyanin and S 35 -azo-bovine-serum-albumin, except the hemocyanin was retained to a greater extent than the albumin. There was very little if any correlation between the number of secondary injections and retention of the initial injection. Quantitative antibody nitrogen data, obtained for the serum of each rabbit showed, in general, an inverse relationship between circulating antibody and radioactivity retained, i.e . the higher the circulating antibody titer, the lower the retention of radioactivity in liver tissue. Passively administered homologous antibody did not produce a change in the retention of the primary injection of antigen nor did secondary injections of a heterologous native protein injected according to the same immunization schedule as the homologous azoprotein. From these results it may be concluded that an intracellular antibody-forming activity influences the loss (or retention) of antigen deposited in liver tissue and that the mechanism is immunologically specific. Footnotes Submitted: 2 October 1957

Barry, Guy T.

doi: 10.1084/jem.107.4.507pmid: 13513915

All acidic carbohydrate has been isolated from the culture medium of the enteric microorganism E. coli K 235 L + O. This substance has been named colominic acid. N -acetylneuraminic acid, which has been isolated from the hydrolysate of colominic acid, is believed to be the monomer unit from which colominic acid is constituted. Colominic acid is not antigenic in rabbits. Tests have failed to reveal a relationship between the ability of E. coli K 235 to elaborate colominic acid and its virulence in mice. Footnotes Submitted: 10 December 1957

Boehme, Diethelm; Dubos, René J.

doi: 10.1084/jem.107.4.523pmid: 13513916

Administration of small amounts of salmonella endotoxin (20 µg.) or of acetone-extracted BCG cells (100 µg.) increases the resistance of mice to infection with Myco. fortuitum as well as their ability to clear carbon particles from their blood stream. Whereas the increased resistance to infection persists for many weeks, the clearing power returns to a normal level within a few days. When normal mice are infected with Myco. fortuitum , there occurs during the first phase of the infectious process a rise in clearing power for carbon particles followed by a fall during the terminal phase of the disease. The rise occurs more rapidly and is more pronounced in animals previously treated with salmonella endotoxin or with killed BCG cells. This acceleration and intensification of the phagocytic response to infection can be detected even in animals which exhibit a normal phagocytic index when tested several weeks after administration of endotoxin or of BCG. Although increase in resistance to infection is correlated with activation of the so called reticulo-endothelial system, there is no evidence of any direct causal relationship between the two phenomena. Footnotes Submitted: 2 December 1957

Stevens, Kingsley M.; McKenna, John M.

doi: 10.1084/jem.107.4.537pmid: 13513917

Ten µg. of the lipopolysaccharide endotoxin of Salmonella typhosa was given to rabbits intravenously to enhance the subsequent antibody response to an unrelated substance. The spleens were removed 24 hours later, diced, and incubated 1 hour with the antigen, bovine-γ-globulin (BGG), in a protein-free medium. After washing, the tissues either were extracted at once or planted and the fluids and tissues harvested 1 to 3 days later. Antibody was determined by a modification of the Boyden hemagglutination technique. Small amounts of antibody were synthesized as early as 1 hour after the addition of antigen. The antibody formed could be specifically inhibited with BGG, was not dialyzable, and did not sediment at 105,000 g for 2 hours. Dose-response studies revealed no antibody formation when the BGG concentration was 0.005 or 0.05 mg./ml. The best responses were obtained at concentrations of 0.5 to 5.0 mg./ml. These results were found irrespective of whether the animal had previously received BGG in vivo . Forty per cent autologous serum increased antibody formation about 9-fold over that secured with protein-free medium or with 40 per cent homologous serum. Antibody formed with this system could be detected by 50 per cent complement fixation test, although at much lower titer than found by hemagglutination. While spleens from rabbits previously given BGG did not produce more antibody than spleens from normal rabbits, they differed in that they produced antibody without the involvement of endotoxin. Under appropriate circumstances, endotoxin was effective in vitro in enabling spleen fragments to produce antibody to BGG. Cortisone acetate administered to rabbits prior to the removal of the spleen severely inhibited antibody production in vitro . Sodium prednisolone phosphate added in vitro showed a similar irreversible effect at concentrations as low as 2 x 10 –5 M . Nitrogen mustard inhibited antibody formation at concentrations as low as 10 –4 M . Footnotes Submitted: 21 November 1957

Rogers, David E.; Melly, Marian Ann

doi: 10.1084/jem.107.4.561pmid: 13513918

Rabbits given an intravenous injection of an 18 to 24 hour broth culture of E. coli commonly died within 28 hours. The injection of a 4 hour broth culture of the same strain of E. coli containing equal numbers of living bacilli produced only an occasional death. Initial clearance rates and subsequent bacteriemias were similar in animals receiving either culture. Study of changes in circulating leukocytes or the temperature response to washed bacterial cells or culture filtrates failed to reveal obvious differences in host response to young or old cultures. It was found that both living bacterial cells and some substance or substances present in culture filtrates were required to produce subsequent death. The injection of whole old cultures containing both these factors produced hypothermia instead of the endotoxin type fever response which followed the injection of whole young cultures. Subsequent experiments revealed that this hypothermia appeared to be secondary to a period of transient but profound shock which occurred soon after the injection in rabbits receiving old cultures. No significant alterations in arterial pressure accompanied injections of young cultures. Evidence is presented which suggests that this period of hypotension rendered animals more likely to die with a persisting bacteriemia tolerated without event by non-shocked animals. The mechanisms which operate to increase mortality during the post shock period are as yet unclarified. Footnotes Submitted: 25 December 1957

Deming, Q. B.; Mosbach, E. H.; Bevans, M.; Daly, M. M.; Abell, L. L.; Martin, E.; Brun, L. M.; Halpern, E.; Kaplan, R.

doi: 10.1084/jem.107.4.581pmid: 13513919

Rats on a stock diet with added cholesterol, cholic acid, and thiouracil developed increased concentrations of cholesterol, total lipide, and beta lipoprotein in the serum, and an increased content of cholesterol in the liver and carcass, despite the fact that the diet produced a cessation of endogenous cholesterol synthesis. Rats with high serum lipide concentrations developed intimal lesions similar to those of human atherosclerosis. The induction of hypertension by desoxycorticosterone and salt accelerated the development of hypercholesterolemia, hyperlipemia, increase in tissue cholesterol content, and atherosclerotic changes in the intima. Hypertension induced by renal artery constriction also intensified the hypercholesterolemia and hyperlipemia. On the other hand, rats receiving desoxycorticosterone acetate without salt or salt without desoxycorticosterone acetate did not show any intensification of hypercholesterolemia or hyperlipemia. The extent of the atherosclerotic lesions was correlated with the concentration of cholesterol in the serum. There was also a positive correlation between blood pressure and the degree of hypercholesterolemia. It remained uncertain whether the increase in atherosclerosis in the hypertensive animals was dependent on the increased lipide content of serum and tissues or on a local effect of the elevated blood pressure. Footnotes Submitted: 16 October 1957

Kleinfeld, Ruth; Melnick, Joseph L.

doi: 10.1084/jem.107.4.599pmid: 13513920

A cytological study was made of short term primary cultures of monkey kidney epithelial cells grown in serum-containing (M-H) and synthetic (SM-2) media. The mitotic activity of the cultures reached a peak 3 to 5 days after seeding, and rapidly declined if the medium was not changed. With replenishment at 48 hour intervals a high mitotic rate was maintained through the 8th day. Cultures subjected to a glycine- and glucose-deficient medium showed a sharp decline in mitotic activity within 24 hours. When such cultures were replenished a burst of mitotic activity occurred 20 to 30 hours later. Mitotic aberrations such as multipolar spindles, anomalous spindles with scattered chromosomes, lagging or precocious movement of chromosomes, chromosome fragments, and chromosome bridges were found in all cultures ranging from 16 to 40 per cent of the dividing cells. Non-mitotic aberrations such as giant nuclei, multinucleate cells, nuclear fragmentation, micronuclei, nuclear inclusions, and phagocytosis were found in approximately 3 to 7 per cent of the non-dividing cell population. The frequency and types of mitotic abnormalities were generally independent of the medium used and the mitotic index of the cultures. Footnotes Submitted: 16 December 1957