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Biomedical subjects

G Henle

Publications and source records attributed to G Henle.

At least 181 records · Page 10Linked to original sources

Relation between Epstein-- Barr viral and cell membrane immunofluorescence in Burkitt tumor cells. II. Comparison of cells and sera from patients with Burkitt's lymphoma and infectious mononucleosis.

Blastoid cell cultures derived from leukocytes of patients in the acute stage of infectious mononucleosis (IM) and harboring Epstein-Barr (EB) virus in at least 1% of the cells were found to possess antigens in their membranes which presently are indistinguishable from those detected in Burkitt's lymphoma (BL) cells by the techniques employed. It was noted that, in the course of IM, antibodies are formed which react in indirect immunofluorescence tests with membrane antigens of live cells from Burkitt tumor lines as well as from IM leukocyte cultures, including an autochthonous line in the case of one patient. Sets of sera from IM patients were tested which included a serum collected weeks to years before onset of illness. In the majority of these the pre-IM serum failed to react in membrane immunofluorescence (MIF) tests with any of several Burkitt tumor cell lines employed, but in some cases the presera reacted with cells of some but not others of the lines. Possible explanations for these discrepant results have been discussed. The antibodies involved in the MIF test are evidently distinct from those responsible for the EBV and heterophile reactions. Maximal MIF activity is attained long after the other two antibodies have reached maximal titers and antibodies to EBV and membrane antigens seem to persist for years whereas the heterophile reaction turns negative within a few months.

Antibodies↗

Effect of arginine-deficient media on the herpes-type virus associated with cultured Burkitt tumor cells.

After incubation at 37 C for 2 or more days, Basal Medium Eagle (BME) supplemented with 25% fetal calf serum (FCS) was not able to support adequate growth of EB3 and other lines of Burkitt tumor cells. The medium did, however, increase, by a factor of about 10, the number of cells synthesizing herpes-type virus with which the cultures were persistently infected. Not every lot of FCS produced these effects, nor were these effects seen when BME and FCS were incubated separately for 7 days before the medium was completed. At 37 C, appropriate lots of FCS interacted with several of the amino acids present in BME; this interaction resulted in an inhibition of cellular growth, whereas interaction with arginine yielded the virus-enhancing effect. Arginine-free BME, supplemented with 25% FCS and used without prior incubation, prevented cellular replication and promoted viral infection to a similar extent as did preincubated complete medium. Replenishment of arginine reduced, but did not regularly abolish, the virus-enhancing activity of preincubated media. RPMI-1629 medium was less affected by preincubation with FCS because it contained twice the amount of arginine that BME contained. The FCS factors which act upon arginine and other amino acids are not dialyzable and are partially resistant to heating at 56 or 60 C for 30 to 60 min. Calf and horse sera appear to be devoid of these activities. The nature of these interactions, as well as the mechanism by which arginine deficiency enhanced the viral infection, remains to be ascertained.

Animals↗

Demonstration of a herpes group virus in cultures of peripheral leukocytes from patients with infectious mononucleosis.

CULTURES WERE INITIATED WITH PERIPHERAL LEUKOCYTES FROM INDIVIDUALS: (i) in the acute stage of infectious mononucleosis (IM); (ii) with past histories of IM; and (iii) without histories of IM and without antibodies to EB virus (EBV). In confirmation of other reports, the first group of cultures developed readily and rapidly into lines of blastoid cells. Cellular replication commenced in 24 of 25 attempts within 17 to 28 days, regardless of the technique employed; i.e., initial seeding of the leukocytes with or without addition of phytohemagglutinin onto monolayers of human diploid cells (WI-38) or direct establishment of suspension cultures. EBV was detected in all cultures by immunofluorescence and also by electron microscopy when tested. Furthermore, the nine cultures which were examined cytogenetically revealed the C-group (#10) chromosomal marker previously found in cultured Burkitt tumor cells. These findings supported the earlier conclusion that EBV is related to, if not identical with, the causative agent of IM. Cultures of the second group commenced growth only within 30 to 60 days in five of seven attempts, depending apparently upon the early presence of WI-38 cells. These cultures also revealed the presence of EBV and, in the three examined, the C-group chromosomal marker. Leukocytes of the third group, seeded onto WI-38 monolayers, failed to become established in four attempts. The possible implications of these findings are discussed.

Culture Techniques↗

Herpes-type virus and chromosome marker in normal leukocytes after growth with irradiated Burkitt cells.

Cultured cells derived from male patients with Burkitt's lymphoma and harboring herpes-type virus particles were lethally irradiated. These irradiated cells induced normal peripheral leukocytes of female infants to grow within 2 to 4 weeks after mixed cultivation. Cells of a line free of this agent failed to stimulate growth. If either type of cell was cultured separately, it did not survive under the experimental conditions. Herpes-type viral antigen and C-group chromosomal marker previously described in cultured Burkitt cells were found in all of the female cell cultures that were obtained.

Burkitt Lymphoma↗

Comparative study of cultured Burkitt tumor cells by immunofluorescence, autoradiography, and electron microscopy.

Cultured Burkitt cells were examined by immunofluorescence, autoradiography, and electron microscopy in an effort to identify the stainable cells with those harboring herpes-type virus particles. Immediately after a 2-hr pulse of (3)H-thymidine, from 30 to 60% of the cells revealed heavy nuclear labeling. In most cases the grains were evenly dispersed, but in about 3 to 5% the grains showed a focal distribution and occasionally they extended into the cytoplasm. Such nuclear foci were rarely seen at 8 hr after the pulse. When the analysis was restricted to preselected immunofluorescent cells, up to 80% showed label at 8 hr and cytoplasmic grains were prominent. To reduce cellular deoxyribonucleic acid (DNA) synthesis, cells were X-irradiated with 3,000 to 6,000 R, and the isotope pulse was applied 1, 4, or 7 days later. Whereas the total number of labeled cells decreased in roughly twofold steps at the respective intervals (from 40 to 10%), the incorporation of (3)H-thymidine into fluorescent cells was not affected by X irradiation. In each series, about 70% of the fluorescent cells contained label when they were examined at 24 and 48 hr after the pulse, whereas at 8 and 72 hr fewer were positive. At the earlier intervals, unlabeled fluorescent cells most likely represented cells which had completed viral DNA synthesis prior to the pulse; at the later intervals, unlabeled fluorescent cells were probably cells which commenced viral replication after the pulse. These data support the conclusion that the immunofluorescent cells are the ones which harbor virus, and also confirm the expectation that the virus is a DNA virus from a member of the herpes group. This conclusion was firmly established by sectioning and electron microscopic examination of individual fluorescent cells, all of which contained numerous virus particles, whereas the nonstained cells prepared in a similar manner were free of them.

Autoradiography↗

Immunofluorescence in cells derived from Burkitt's lymphoma.

Henle, Gertrude (Children's Hospital of Philadelphia, Philadelphia, Pa.), and Werner Henle. Immunofluorescence in cells derived from Burkitt's lymphoma. J. Bacteriol. 91:1248-1256. 1966.-Indirect immunofluorescence tests led to the brilliant staining of a small proportion of the cells in five different cultures derived from Burkitt's (African) lymphomas. The reaction was not restricted to the 17 sera from cases of this disease but extended to many sera from American individuals, whether healthy donors or patients suffering from a variety of illnesses. The incidence of positive sera increased with age from about 30% in childhood to > 90% in adults. Fluorescein-isothiocyanate-conjugated human gamma-globulins were suitable for direct staining of the same proportion of cells. The stained cells appeared to be in varying stages of degeneration, but cultural conditions leading to an increase in the cellular death rates failed to result in a rise in fluorescent cells. Several observations suggest that the stainable cells might be those which are seen to harbor virus particles under the electron microscope. Two cell lines derived from leukemic patients in this country also contained a small fraction of stainable cells but two others, and numerous primary human leukocyte cultures, gave consistently negative results. Attempts to relate the staining to known viral antigens have failed to implicate herpes simplex, varicella, cytomegalo, and reo viruses types 1, 2, and 3. The nature of the virus carried by the lymphoma cells as well as of the staining reactions remains to be determined.

Adult↗

Complement-fixation tests with cell lines derived from Burkitt's lymphoma and acute leukemias.

Armstrong, Donald (Children's Hospital of Philadelphia, Philadelphia, Pa.), Gertrude Henle, and Werner Henle. Complement-fixation tests with cell lines derived from Burkitt's lymphoma and acute leukemias. J. Bacteriol. 91:1257-1262. 1966.-Cells of various lines isolated from Burkitt's lymphomas and acute leukemias and disintegrated by freezing and thawing or sonic treatment were found to react in complement-fixation tests with a considerable proportion of human sera. At least 10(7) cells per milliliter were required for antigenic activity. All but one of 13 sera from Burkitt lymphoma patients were positive, with titers ranging from 1:8 to 1:320. About 20% of sera from American children and 60% of sera from adults, regardless of diagnosis, showed titers in a similar range. Sera giving positive tests with one of the neoplastic white cell antigens usually reacted also with many if not all of the others, but rarely with antigens derived from normal peripheral leukocyte cultures and not at all with HeLa or other human nonleukocytic cells. Various observations indicate that the complement-fixation test measures mainly antigens which are different from those detected by immunofluorescence. The nature of the reactions described remains obscure.

Adenocarcinoma↗