Enhancement of Lewis lung carcinoma by the concomitant infection of the host with herpes simplex virus type 1 and type 2.

Infection with herpes simplex virus (HSV) enhances the growth of Lewis lung tumor (LLT). Infection with HSV-1 enhances as efficiently as HSV-2. A significant acceleration of primary tumor formation was obtained. In mice, lung metastases were increased in number and size above the controls. Enhancement occurred only with high doses of infectious virus and was also obtained only when the virus infection was at a site close to that of the tumor implantation. This finding, in addition to the lack of antigenicity of LLT, is interpreted as arguing against an immunologic mechanism of the phenomenon.

Augmented immunogenicity of Lewis lung carcinoma by infection with herpes simplex virus type 2.

In vitro, LLT cells sustain HSV-2 replication without evidence of lysis. Simultaneously, multiplication of the cells is stimulated. These xenogenized cells were tested for their immunopotentiating capacity: three-step immunization with xenogenized viable cells conferred significantly augmented transplantation resistance to a challenge graft with 4 x 10(4) intact LLT cells. Latency periods preceding tumor formation were increased and 15% of the mice failed to form primary tumors. Metastasis was likewise decreased and 25% of the mice had healthy lungs. Immunopotentiation, however, did not suffice to significantly protect against a challenge with 6 x 10(4) intact cells. The presence of virus-specific neoantigens on HSV-2-infected viable cells was demonstrated by the progressively increasing number of rejections of 4 x 10(4) xenogenized cells during the successive immunization steps. Immunization with non-viable LLT cells did not augment resistance to challenge.

Isolation and characterization of the growth-stimulating factor in supernatants of herpes simplex virus type 2 infected cells.

Lytic infection of CV1 cells with herpes simplex virus type 2 does not stimulate ornithine decarboxylase activity and there is no correlation between polyamines and a growth-stimulating factor (GSF) which is present in the supernatant of the cultures. The factor was partially purified by gel filtration on Sephadex G-50 and polyacrylamide gel electrophoresis. Gel filtration as well as dialysis through membranes with different pore diameters indicated a molecular weight between 3,500 and 10,000 daltons. GSF is not extracted from aqueous solutions by diethyl ether. The activity is partially sensitive to trypsin digestion and is completely destroyed by periodate oxidation. These results suggest that GSF is a glycoprotein or a glycopeptide whose mitogenic activity resides mainly in the sugar moiety.

Herpes virus infection as a cofactor in carcinogenesis: supernatants of herpes virus type 1- and type 2-infected cell cultures contain a cell growth-stimulating factor.

Some cell cultures synthesize a mitogenic factor (DNASF) following their lytic infection with either HSV-1 or HSV-2 which is shed into the medium. Of six cell lines permissive to HSV infection tested, three produce DNASF (CV1, HF, MCA) and a significantly higher (3H)-TdR incorporation was obtained with indicator cells growing in supernatants of virus-infected cells (SupV+) than in the corresponding supernatants of sham-treated cells (SupV-). Sometimes the values obtained with SupV+ exceed those obtained with cell controls, which demonstrates that DNASF is not simply a nutritional factor. Other cell lines do not produce DNASF (HeLa, Wistar and probably REF) and SupV+ frequently inhibit cell growth. DNASF is genetically nonspecific and all indicator cells tested were stimulated by supernatants which contain the mitogen, including the nonpermissive DC-3F cells. The quantity of DNASF induced is directly proportional to the number of cells in the culture and its production peaks when the lysis of the cells is complete. It is not accumulated inside the cells.

[Herpes simplex virus (hsv-1 and hsv-2) infection: its clinical and oncogenic properties (author's transl)]. / Die Herpes-simplex-Virus-(HSV-1-und HSV-2-) Infektion- ihre klinische und onkogene Bedeutung.

The epidemiology and clinical features of diseases caused by the herpes simplex virus, are reviewed and recent results are discussed which give an insight into the complex mechanism of primary and chronic, recurrent HSV-1 infections. Immunological reactions in HSV infection and data concerning the oncogenic potential of HSV-1 and -2 are dealt with. Furthermore, current therapeutic possibilities are outlined.

An investigation into the relationship of the immunological status of Herpes simplex virus type-2 (HSV-2) infected animals and the tumor-enhancing property of this infection.

In order to compare the state of immunity of HSV-2 infected animals with their sham treated counterparts, several in vivo methods for cell-mediated immunity in addition to the stimulation in vitro of mouse splenocytes with PHA were performed. The grafting of C 3 H mice with heterologous skin and the paw test with PHA showed no noticeable difference between the two groups. The tuberculin test performed on guinea pigs infected via the footpads, permitted the detection of a slightly impaired reactivity of the virus-infected animals. The PHA assay in vitro showed reduced stimulation indices with cultures of splenocytes from virus-infected mice about 10 days p.i. These reduced stimulation indices are the result of 1. a low 3 HTdR incorporation by the PHA stimulated cultures and 2. of a relatively high 3 HTdR uptake by the unstimulated control cultures. An interpretation of these results is suggested. Finally, the possible implications of a shortlived impaired cell-mediated immunity on the tumour enhancing property of the HSV-2 infection, which has been described in previous publications, are discussed.

The phosphorylation of histones in herpes virus infected Vervet-monkey kidney cells (CV-1).

The phosphorylation of histones of the F1 group and of fraction F2a2 is stimulated in monkey kidney cells (CV-1) to almost two times the control values 14 hours after their infection with herpes virus, type 2. At the same time high amounts of viral DNA are produced. It seems very likely that the stimulated phosphorylation of these histone fractions is a prerequisite for the enhanced synthesis of DNA and possibly also RNA.

[Investigations concerning the evidence of herpes simplex virus type 2 (HSV-2) antibodies in patients with carcinoma of the cervix (author's transl)]. / Untersuchungen zum Nachweis von Herpes-simplex-Virus-Typ-2-(HSV-2-)Antikörpern bei Zervixharzinompatientinnen

The neutralizing antibodies to HSV-1 and HSV-2 were determined in the sera of 128 patients. Infection was detectable in nearly 100% of the cases in each of the three investigated groups (patients with carcinoma of the cervix, female patients with chronic recurrent HSV infection in the genital area and a control group without and history of HSV infection). The percentage of patients displaying HSV-2 antibodies in the group with carcinoma of the cervix (38%) is significantly higher than in the control group (12%). The results are compared with the findings of other authors and the possible causal significance of HSV in carcinogenesis is discussed.

The tumor enhancing property of herpes simplex virus type-2 (HSV-2).

The s.c. infection of DBF-1 mice with HSV-2 has a tumor enhancing effect on simultaneously i.m. implanted MCA-induced syngeneic spindle cell sarcoma. Thus with the dosage used here, the first palpable tumors appeared on day 6 in the virus treated group as compared to day 9 p.i. in the sham treated batch. Further, more tumors were formed: 75% among the infected and 40% among the sham injected. The tumor yield could be modulated: no difference between the 2 groups was found when the tumor cell dose was increased sufficiently to advance the first appearance of tumors to less than 6 days. The same result was obtained when the neoplastic cells were implanted 3 days ahead of injection of the virus. When the dose of the cells was decreased, no tumors were found in the sham treated batch, whereas there were some in the infected group. Previous in vitro mixing of the neoplastic cells with virus completely prevented tumor formation, probably due to their lysis by the virus. In fact, in tissue cultures, MCA cells become lytically infected with HSV-2. It is speculated that a similar situation occurs in acutely ill mice which die before the 13th day p.i., i.e., the tumor cells are destroyed by the virus. In fact, those animals, as a rule are free of neoplasmas. Thus it appears that an acute HSV-2 infection prevents MCA sarcoma whereas a latent infection promotes its growth. The possible reasons for this tumor enhancement caused by HSV-2 infection are discussed.

The soluble antigens of Rickettsia prowazeki, R. typhi and R. canada. Investigation of their interrelationship by various serological methods.

The purpose of this research is the isolation of an eventual species-specific fraction from the "soluble antigen" of Rickettsiae. The "soluble antigen" of R. prowazeki (Breinl strain), R. typhi (Wilmington strain) and R. canada were purified at 25% saturation with ammonium sulphate (PSA). Corresponding antisera were produced in rabbits. The serological methods used were the complement fixation, the micro-agglutination, the precipitation method in capillary tubes and the immuno-diffusion method carried out with complete and previously cross-absorbed antisera. Furthermore, the PSA were subjected to immuno-electrophoretic and disc electrophoretic fractionation. The PSA of R. prowazeki was found to contain at least 4 group-specific fractions. A species-specific component could be demonstrated with certainty only with the precipitation method in capillary tubes carried out with previously cross-absorbed antisera. The PSA of R. typhi contains 5 fractions of which 4 are group-specific and one is species-specific. This result was confirmed by all methods. The PSA of R. canada: the maximum of 3 components could be detected with the help of immuno-electrophoretic fractionation. A fourth antigenic determinant was made apparent by the presence of corresponding antibodies in the anti-R. canada PSA only.

A study of the immuno-suppressive activity of Herpes simplex virus type 2 and the tumor enhancing potential of the virus on Yoshida sarcoma.

The relative delays of Yoshida sarcoma (YS) tumor induction were used as indicator for the immunosuppressive potential caused by the subcutaneous infection of Sprague-Dawley rats with approximately 10(7) TCD50 HSV-2. The tumor formation is clearly accelerated and the number of tumors is increased as compared to sham injected rats. The impairment of immunity is at its maximum when the virus and the YS cells are applied simultaneously, whereas virus given after the tumor is of no consequence. The adoptive neutralization test of Winn showed that the neutralization potential of spleen cells originating from HSV-2 infected donors is decreased from day 8 to day 11 post infection but rturns back to normal a week later. The stimulation of spleen cells with PHA is reduced up to 50% of control values on day 6 to 8 post virus infection but PFC and RFC values are not noticeably affected and neither are the hemagglutination titres. The implications of the immuno-suppressive faculty of HSV-2 for human genital malignancies are discuss.ed.