Experiences in the search for anti-inflammatory agents of microbial origin.

Whole shaken cultures of 20 random, unidentified actinomycetes were extracted with n-butanol at pH 4.5, 7.0 and 8.5, respectively. Residues of butanol-extractable materials (BXM) were reconstituted (100X) in buffers and freeze-dried. BXM were surprisingly well tolerated in animals and were screened against influenza A viral pneumonia in mice. One culture yielded BXM-80 which suppressed both chemical (LPS) and viral (NDV) pneumonia in mice as well as inhibited rat foot pad edema induced by carrageenin. Aspirin, Butazolidin, hydrocortisone, indomethacin, and prednisolone, which are known to inhibit carrageenin-induced rat foot pad edema were tested against chemical (LPS) and viral (NDV) pneumonia in mice. Only hydrocortisone and prednisolone suppressed LPS pneumonia. All of these 5 compounds failed to inhibit NDV pneumonia. Microbial products are suggested as a source for new and unique anti-inflammatory agents.

Responses of isolator-derived Japanese quail and quail cell cultures to selected animal viruses.

Thirteen oncogenic and necrotizing animal viruses were assayed in LIFE Sciences, Inc. (LSI)-specific pathogen-free Japanese quail and LSI-specific pathogen-free chicken embryo cell cultures. Nine viruses produced similar titers in the quail and chicken cell systems, whereas four viruses showed significantly higher titers in chickens. Young Japanese quail and chickens were inoculated with five selected avain viruses and maintained in stainless-steel isolators. Comparable responses were noted in quail and chickens injected with Newcastle disease virus and avain leukosis virus, but quail were significantly more resistant than chickens to fowl pox virus, laryngotracheitis virus, and Marek's disease herpesvirus. Although no overt symptoms of disease were observed in Japanese quail inoculated with most avain viruses, neutralizing antibody or virus was detected, indicating presence of an inapparent infection. In one experiment, neutralizing antibody was detected in a comparable number of quail and chickens after inoculation with avian leukosis virus. Avian leukosis virus viremia was observed at 12 and 70 days postinoculation, with the COFAL (complement fixation for avian leukosis) titers similar for quail and chickens. Most quail infected with Marek's disease herpesvirus produced neutralizing antibody within 70 days but showed no classical symptoms of Marek's disease even when held for 5 months. In contrast, all chickens inoculated with Marek's disease herpesvirus died within 20 days. The utility of quail embryo cell cultured in the preparation of vaccines and biological reagents is discussed.

Biological and biochemical evidence for an interaction between Marek's disease herpesvirus and avian leukosis virus in vivo.

The DNA-containing Epstein-Barr herpesvirus has been implicated in the etiology of Burkitt's lymphoma, a malignant tumor of children in Africa. Recently, however, particles possessing four biochemical characteristics of RNA tumor viruses have also been identified in these tumors. The fact that both types of viruses are found suggests that an interaction between them may be playing a role in the etiology of Burkitt's lymphoma. To explore this possibility with a defined animal model, experiments were performed with the avian Marek's disease, a malignant lymphoproliferative disease associated with a herpesvirus. Controlled contact studies demonstrated that tumorigenesis in a line of isolator-derived, barrier-sustained, specific pathogen-free chickens requires exposure to both the Marek's disease herpesvirus and an avian leukosis virus, Rous-associated virus, type 2. Exposure to either agent alone did not result in tumors. Molecular hybridization experiments to cytoplasmic RNA from similarly contact-exposed conventional Cornell S-line chickens provided further evidence for the occurrence of an interaction between Marek's disease herpesvirus and the avian leukosis virus.

Growth curve and distribution of Rous sarcoma virus (Bryan) in Japanese quail.

On primary infection with the Bryan strain of Rous sarcoma virus (RSV), the growth curve of the virus in the brain of Japanese quail was similar to that observed in chicks and turkey poults. Infectious virus disappeared from the brain after inoculation. After an eclipse period during which no virus was detectable, infectious virus began to appear at 2 days and reached maximal titers in the brain samples at 7 days after inoculation. When Japanese quail were infected intracerebrally with RSV, relatively high titers of virus were recovered from brain tissue but not from liver, lung, kidney, or blood of moribund birds. Only tumors produced in the wing web of quail infected subcutaneously yielded high titers of virus. Other tissues yielded no virus, even though wing web tumors appeared as early as in chicks similarly infected. RSV could be propagated in the wing web of quail for at least 14 passages without any loss of infectivity. On the other hand, serial passage in quail brain resulted in a progressive loss of infectivity until virus was completely lost.