Dd Mice Research Articles

Experimental Study on the Relapse of Tuberculosis after the Termination of Antituberculous Chemotherapy Using Immunodeficient Nude Mice

The effect of cellular immunity induced by tuberculous infection on bacteriological relapse after the termination of antituberculous chemotherapy was studied experimentally using immunodeficient nu/nu mice and immunocompetent dd mice. The efficacy of intensive chemotherapy was excellent even in nu/nu mice; tubercle bacilli in the organs decreased below the detectable limit, but formidable regrowth of bacilli was seen after the termination of chemotherapy. On the other hand, in the case of dd mice that established antituberculous cellular immunity through tuberculous infection, bacteriological relapse was generally very slight. It was concluded that bacteriological relapse was related closely with the established cellular immunity induced by the infected tubercle bacilli.

Influence of the Ah locus on the humoral immunotoxicity of 2,3,7,8-tetrachlorodibenzo- p-dioxin: Evidence for Ah-receptor-dependent and Ah-receptor-independent mechanisms of immunosuppression

There are conflicting reports in the literature regarding the role of the Ah locus in 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) immunotoxicity. The present studies have utilized two congenic strains of C57Bl 6 mice that differ only at this locus to assess its influence on TCDD-induced suppression of antibody responses. Mice were given a single oral dose of TCDD 2 days prior to challenge with sheep red blood cells (SRBC) or trinitrophenyl-lipopolysaccharide (TNP-LPS). The subsequent dose-dependent effects of TCDD on the amount of antibody produced by splenic plasma cells were measured using the hemolytic antibody isotope release assay. In addition, the relative importance of the Ah genotype of lymphoid versus nonlymphoid tissue was examined in adoptive transfer experiments. Aryl hydrocarbon hydroxylase (AHH) activity was significantly induced in Ah bb mice by a dose of 0.5 μg/kg TCDD and maximally induced by a dose of 2 μg/kg. Ah dd mice required 10-fold higher doses of TCDD to induce comparable levels of AHH. The degree of thymic involution and liver hypertrophy induced by TCDD was also influenced by the Ah genotype of the animals. Both Ah bb and Ah dd mice exhibited dose-dependent suppression of the anti-TNP response following TCDD exposure. The ID50 was 7.0 μg/kg in Ah bb mice and 30.8 μg/kg in Ah dd mice. Suppression of the antibody response to SRBC was also dependent on the Ah locus. The ID50 in Ah bb mice was 0.6 μg/kg TCDD. However, an apparent biphasic dose response for suppression of the anti-SRBC response in Ah dd mice suggested the involvement of an Ah-independent component of suppression as well. In adoptive transfer studies, lymphocytes were identified as an Ah-dependent component of the response. The Ah-independent component of the response was not identified, and could be either lymphoid or nonlymphoid in nature. The possibility that T helper cells represent the Ah-independent component is discussed.

Promoting effect of basic lead acetate administration on the tumorigenesis of lung in N-nitrosodimethylamine-treated mice

Lead compounds are frequent environmental contaminations and some of them have been found to be carcinogenic for animals, although epidemiological studies have not been considered to provide sufficient evidence that exposure to lead or lead compounds causes cancer in humans. There re only a few reports on the promoting effect of lead on chemical carcinogenesis in vivo. By the concurrent administration of lead compounds and certain organic carcinogens to rats or hamsters, the cocarcinogenic activity of lead has been found in the kidney of rats or the lung of hamsters. In this report the promoting effect of posttreatment with basic lead acetate (BLA) on the development of lung tumors in strain dd mice exposed to N-nitrosodimethylamine (NDMA) was examined. The concentration of lead and the activity of {gamma}-glutamyltranspeptidase ({gamma}-GTP) in the lung during the carcinogenicity experiment were also measured.

ドイツマウスの導入に関する考察―秦佐八郎の関わり―

An inbred strain of mice, DDD, was established at the Institute for Infectious Diseases (so-called Denken in Japanese; currently called Institute of Medical Science, The University of Tokyo: Jms) in 1962. Many interesting characteristics including development of pregnancy-dependent mammary tumors, presence of Fv-2s, a Friend leukemia virus-susceptible gene, high incidence of hydronephrosis and low level of natural killer (NK) cell activity have been reported in this strain of mice. Two congenic strains, DDD-Fv-2r and DDD-Mtv-2+, have been produced. Inbreeding of dd mice maintained at Denken was commenced in 1957 and the resulting inbred strain was named DDD after dd at Denken. The ancestors of dd mice had come from Germany. The process of introduction to Japan and the history of these mice were surveyed in the literatures, since the description concerning them had been confused. As a result, the following history was confirmed: the original colony of mice of an unknown size was introduced from the Hoechst Company, Frankfurt a.m. Main, Germany into the Kitasato Institute, Tokyo, Japan by Dr. Tsuneo Komatsu under the direction of Dr. Sahachiro Hata in autumn, 1928. Dr. Hata and his colleagues bred these mice to use for medical researches and called them "deutsche Maus". Their descendants (one male and one female) were shipped to the Health Institute of Manchuria Railway, Talien (currently called Dalian), China on demand of Dr. Koji Ando in 1934. Two males and eight females from the Talien colony were shipped back to Dr. Saburo Kojima, the Institute for Infectious Diseases, Tokyo, Japan in 1943. They had been maintained without crossing with another colony at the Institute, from which mice were distributed to many institutes throughout Japan between 1944 and 1960, and served as the starting nuclei of many inbred or outbred strains. The "deutsche Maus" was named dd mice after "deutsche Maus at Denken" in 1952. Thus, the mouse strains connected with dd mice usually have D, DD or dd at the head in their names.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-induced lipid peroxidation in genetically responsive and non-responsive mice

The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) administration on lipid peroxidation in various strains of responsive and non-responsive mice was determined. The hepatic content of thiobarbituric acid reactive substances (TBARS) was used as an index of lipid peroxidation, employing malondialdehyde as the standard. Six days after the administration of a single oral dose of 0.5 μg TCDD/kg to congenic male C57BL/6J mice, which were either homozygous (bb) or heterozygous (bd) with respect to the gene for the Ah locus, significant increases in hepatic lipid peroxidation were induced. Hepatic lipid peroxidation was induced in homozygous non-responsive (dd) mice by a dose of 25 μg TCDD /kg but not 0.5 μg/kg. In kidney, heart and testis, lower doses of TCDD produced lipid peroxidation in C57BL/6J bb and bd mice as compared to dd mice. The ability of TCDD to induce lipid peroxidation in B6D2F1/J, DBA2J and Swiss-Webster mice was also examined. A dose of 2,500 μg TCDD/kg produced the same level of lipid peroxidation in DBA/2J mice that was observed with 180 μg TCDD/kg in C57BL/6J bb and B6D2F1/J mice. Lipid peroxidation was not induced in kidney, heart, or testis at any of the doses of TCDD that were administered to DBA/2J mice. Thus, large strain differences exist in the ability of TCDD to induce lipid peroxidation in mice. In the liver, the ability to induce lipid peroxidation by TCDD is controlled in part by the Ah “b” allele, although other loci may play a major role.

Strain-specific occurrence of two ornithine decarboxylase species in mouse kidney

Ornithine decarboxylase in the crude extract from the kidney of androgen-treated mice was labeled by reaction with radioactive α-difluoromethylornithine and analyzed by sodium dodecylsulfatepolyacrylamide gel electrophoresis followed by autoradiography. Two species of ornithine decarboxylase with molecular weights of 54,000 and 52,000 were observed in the crude extracts from BALB, C57BL, C58, SJL, and dd mice but only the species with a molecular weight of 54,000 was observed in the crude extracts from AKR and C3H mice, indicating strain-specific occurrence of at least two species of mouse ornithine decarboxylase.

The distribution of Langerhans cells in the dorsal mucosa of the mouse tongue.

Ultrastructure and distribution of Langerhans cells in the dorsal mucosa of mouse tongue were observed by light and electron microscopy and by ATPase histochemistry. The epithelium could be divided into papillary and interpapillary parts. Both papillae and interpapillae were orthokeratinized. The differences between the two parts were mainly due to the cellular characteristics of the prickle cell layer, granular layer and keratinized layer. In the interpapillae, tonofilaments in the cytoplasm of the prickle cells evenly dispersed without forming bundles, and the desmosomes were observed more clearly. The granular layer was less developed than that of the papillae. The cytoplasm of the keratinized layer stained with toluidine blue more lightly than that of papillae. By light microscopy, cells with clear cytoplasm occurred in the prickle cell layer just above the basal cell layer of interpapillae. They extended long cytoplasmic processes vertically, up to the granular layer and down to the basal layer. ATPase-positive dendritic cells were distributed in the interpapillae. By electron microscopy, Birbeck granule-containing Langerhans cells were observed in the interpapillae. The dendritic cells, which were observed as cells with clear cytoplasm by light microscopy or as ATPase-positive cells, agreed in morphology and distribution with the Langerhans cells which were observed by electron microscopy. In dd mice, ATPase-positive Langerhans cells were not observed at 2 weeks after birth, whereas they were observed at 3 and 4 weeks.

A transgenic class I antigen is recognized as self and functions as a restriction element.

The function of a transgenic Dd class I molecule in the induction of immunologic tolerance to major histocompatibility complex antigens and in directing major histocompatibility complex restriction in C57BL/6 mice were investigated. All of the transgenic Dd mouse strains were found to be tolerant for the Dd antigen. Spleen cells from transgenic mice were immunocompetent but consistently failed to generate an anti-Dd cytotoxic T lymphocyte response in vitro, and skin grafts between transgenic Dd mice were not rejected. These data suggests that the Dd antigen was recognized as a self molecule. In addition, the transgenic Dd mice generated antigen-specific Dd-restricted cytotoxic T lymphocyte, indicating that the Dd antigen also functioned as a restriction element for antigen recognition. These observations demonstrate the usefulness of the transgenic mouse system for studying class I antigen expression and function.

Strain differences in the reverse tolerance to methamphetamine and changes in catecholaminergic neurons in mice.

The effect of methamphetamine (MAP) on ambulatory activity and neurochemical changes in catecholaminergic neurons in the brain were investigated in dd and C57BL/6 strains of male mice. The mice were given repeatedly MAP at 2 mg/kg, s.c., 10 times at a fixed interval of 4 days. Single MAP markedly increased the ambulatory activity in both strains of mice. The ambulation-increasing effect was progressively enhanced without accompanying stereotyped behaviors when the drug was repeatedly given. The dd mice showed higher susceptibility not only to single MAP but also to the enhancing effect of the drug (reverse tolerance) that the C57BL/6 mice. On the other hand, non-treated dd mice exhibited lower maximum densities of 3H-spiperone binding sites in the striatum and 3H-WB4101 binding sites in the cortex and hippocampus than non-treated C57BL/6 mice. In contrast, the dd mice exhibited higher noradrenaline turnover than the C57BL/6 mice in the brain regions examined. The repeated administration of MAP produced decrease in the densities of both 3H-spiperone and 3H-WB4101 binding sites in the corresponding regions with increase in catecholamine turnover in dd mice. However, the similar changes were observed only in 3H-WB4101 binding sites and noradrenaline turnover in C57BL/6 mice. These results suggest that the ambulation-increasing effect of MAP is positively correlated with catecholamine turnover, while it was correlated negatively with the densities of catecholamine binding sites. Furthermore, the enhancing effect of MAP is supposed to have been partially elicited by changes in brain catecholaminergic systems, in particular an increase in catecholamine turnover.(ABSTRACT TRUNCATED AT 250 WORDS)

Reduced fecundity of Hymenolepis nana due to thymus-dependent immunological responses in mice

Reduced fecundity of Hymenolepis nana due to thymus-dependent immunological responses in mice. International Journal for Parasitology 16: 81–85. The fecundity of the dwarf tapeworm, Hymenolepis nana, was enhanced in congenitally athymic nude CD-1 (ICR) mice but depressed to the same degree as in phenotypically normal littermates when they were reconstituted with thymocytes before infection. The reduction in fecundity of this parasite was clear only when H. nana were recovered from those strains of mice which demonstrate a “late response” against luminal cysticercoid challenge before the established worms become fully mature (within 15 days of initial immunizing egg inoculation). The fecundity of H. nana in dd mice, which acquire the late response within 30–40 days, was greatly advanced than that in BALB/c or CD-1 (ICR) mice and somewhat better than even that in the nude CD-1 (ICR) mice and appeared to be little, or not at all, depressed. The fecundity and longevity of this parasite, highly variable among mouse strains, is discussed in terms of variations in the rapidity of expression of protective immunity against the lumen phase.

Serotonin's influence on predatory behavior of highly aggressive cba and weakly aggressive DD strains of mice

The effects of serotonin were studied on locust-killing behavior of mice from low (DD) and high (CBA) predatory aggressive strains. 5-HTP injected intraperitoneally (50 and 100 mg/kg) or 5-HT administered into the lateral ventricle (10 μg) significantly reduced locust-killing behavior in highly aggressive CBA mice. Imipramine (20, 30, and 40 mg/kg) elicited a dose-dependent inhibitory effect on predatory behavior. Fluoxetine (10 and 20 mg/kg) alone had a slight influence on locust-killing behavior but potentiated the action of the subthreshold dose of 5-HTP (25 mg/kg). Pretreatment with the blocker of 5-HT2 type receptors methysergide (2 mg/kg) abolished the inhibitory effect of 5-HTP. These finding indicate that serotonin of the brain exerts an inhibitory effect on predatory behavior in mice. In contrast, neither lesion of the dorsal raphe nucleus (although significantly depleting the brain serotonin) nor treatment with methysergide (2 mg/kg) induced locust-killing behavior in weakly aggressive DD mice. Low predatory aggressiveness in DD mice is suggested to be related to the low tonus of the mechanisms activating killing behavior rather than to excessive serotonergic inhibitory influences.

Conditioned lever-press avoidance response in mice: acquisition processes and effects of diazepam.

Acquisition of conditioned lever-press avoidance behavior on a continuous avoidance schedule (response-shock interval = 30 s and shock-shock interval = 5 s) and on a discrete avoidance schedule (intertrial interval = 25 s and duration of the warning presentation = 5 s with an escape contingency) in dd mice was investigated. When the behavioral baseline had stabilized, the effects of diazepam on avoidance behaviors were examined. About 60% and 80% of the mice achieved criterion levels of avoidance behavior under continuous avoidance (shock rate being less than 0.5/min) and discrete avoidance (avoidance rate being higher than 75%), respectively. Diazepam (0.5--4 mg/kg SC) dose-dependently impaired avoidance behavior of mice which had a low baseline shock rate and a high baseline avoidance rate under continuous and discrete avoidances, respectively. The changes in avoidance behavior in mice after diazepam were almost identical with those previously found in rats.

Virus-induced immune complex disease: genetic control of C1q binding complexes in the circulation of mice persistently infected with lymphocytic choriomeningitis virus.

Virus-antibody immune complex formation, deposition, and disease is a common manifestation in most mice persistently infected with lymphocytic choriomeningitis virus (LCMV). Although mice of several strains persistently infected with LCMV mount continuous anti-LCMV immune responses to the three virion structural polypeptides, the amount of antibody(s) made varies among strains. The formation of antibody to LCMV correlates with the detection of C1q binding materials (immune complexes) in the circulation; however, there is no correlation between the total amount of IgG and the C1q binding material. SWR/J mice (H2qq) are high responders (high levels of anti-LCMV antibodies, high C1q binding responses), whereas BALB/W mice (H-2dd) are low responders (low levels of anti-LCMV antibodies, low C1q binding responses). Sera from 37 SWR/J mice, 12 wk of age, bound 55.1% of offered 125I C1q compared to 8.4% for 37 age and sex-matched BALB/W mice. SWR/J mice made approximately 60-fold more antibody to LCMV than did BALB/W mice. To define the genetic factors involved, we used the C1q binding assay, high responder and low responder mice, their hybrid offspring, backcrosses of the hybrids to both high and low responder parents, and selected recombinant inbred mouse strains. From studies with BALB/W mice persistently infected with LCMV, we defined low C1q responder mice as those having binding activity of 18.4% or less (BALB/W mean C1q binding value + 2 SD). By using this criteria for 12-wk-old mice, 55/57 (96%) of SWR/J, 27/37 (73%) of F1(SxB or BxS), 13/21 (62%) of qq or 11/20 (55%) qd from F1 x SWR/J, and 6/13 (45%) qd from F1 x BALB/W were high C1q responders. In contrast, none of the 17 dd mice from the F1 x BALB/W cross were high responders. Thus, F1 hybrid mice are high responders (dominant) and data from backcrossing F1 hybrids to either high or low responder parents suggested the complexes associated with C1q binding was controlled by gene(s) in the H-2 complex. Support for the role of Ir gene(s) was provided by experiments showing LCMV persistently infected BALB/kae (H-2 KkIkDk) and recombinant inbred strains B10.A (KkIkDd) and A.TL (KsIkDd) were high C1q responders, whereas B10.A(5R) (KbIbDd) and BALB/cby (KdIdDd) were not. The wide scatter in C1q binding levels observed among C1q high responder mice and the production of C1q binding levels in F1 mice that are intermediate between high and low responder strains suggested that, in addition to H-2-linked genes, other non-H-2-linked genes also play a role in this response. The amount of C1q binding complexes in LCMV persistently infected mouse strains and their crosses was unrelated to the amounts of infectious virus carried in their sera. Despite the low C1q binding by sera of H-2dd mice that originated from mating F1 hybrid (qd) x BALB/WEHI (dd), these mice carried equivalent amounts of infectious virus as their qd littermates or qd and qq mice originated from F1 x SWR/J mice.

Hymenolepis nana: Immunogenicity of a lumen phase of the direct cycle and failure of autoinfection in [formula omitted] mice

When BALB c mice were given BALB c mouse-derived cysticercoids (cysts) of Hymenolepis nana, only 1 43 mice became autoinfected, whereas most ( 31 38 ) of dd mice given the same infection became massively autoinfected with mature worms. When BALB c mice initially given cysts were challenged with eggs on Day 7, just before the patency of the primary infection, there was normal development into cysts, but almost none of them developed into adult worms. Thus, the failure of autoinfection of H. nana in BALB c mice was not a result of failure of eggs to differentiate into cysts in the intestinal tissue, but a result of failure of these cysts to develop into adult worms in the lumen. The reasons why autoinfection does occur in dd and other strains of mice and not in the BALB c strain are discussed in terms of the difference in onset of the late response in these strains of mice, ie., the response that is acquired after egg inoculation, and is directed against the lumen phase of cyst challenges. It is strongly suggested that (1) the lumen phase which follows cyst inoculation is highly immunogenic, but clearly differs from tissue phase which follows egg inoculation, (2) the autoinfection which occurs in some strains of mice is therefore not a result of no or poor immunogenicity of the lumen phase but is due to a delay of onset of the late response with the result that a secondary generation may mature, and (3) in other strains of mice, including BALB c , which acquire the late response within 15 days of initial egg inoculation, autoinfection normally does not occur after cyst infections.

非近交ddマウスにおける乳癌ウイルス (MuMTV) 感染の実態

The expression of mammary tumor virus (MTV) antigen in the milk and various organs of three non-inbred dd mouse stocks (ddO, ddN and ddY) was examined by the immunodiffusion (ID) and micro-immunodiffusion (micro-ID) tests. The rate of MTV antigen expression in the milk was 100% at the first lactation in ddO (6/6) and ddN mice (10/10), and 23% in ddY mice (3/13). Mammary tumor incidence was 13% (mean tumor age: 12.0 months), 32% (9.6 months) and 10% (11.5 months) in ddO, ddN and ddY mice, respectively, In F1 hybrids between MTV-free BALB/c females and dd males, a high level of MTV antigen was detected by the ID test in the milk of (BALB/c X ddO) F1, however, the levels in (BALB/c X ddN) F1 and (BALB/c X ddY) F1 mice were low at the first lactation and elevated with the advance of lactation number. Mammary tumor incidence had a trend to be higher and earlier in these F1 hybrids than in non-inbred dd stocks. The development of mammary tumors and detection of MTV antigen in F1 hybrids indicate the extrachromosomal transmission of MTV by male dd mice. The micro-ID test has shown that the mammary tumors, mammary glands, male genital organs except for the testis and the salivary gland expressed MTV antigen, with a high frequency of suggesting that secondary male genital organs may play an important role in MTV infection in mice.

Thickening of basement membrane of muscle capillary in spontaneously diabetic KK mice.

In order to clarify the relationship between the thickening of muscle capillary basement membrane (MCBM) and diabetes mellitus, an experimental study was carried out using spontaneously diabetic KK mice. Glucose tolerance tests and the measurements of the width of MCBM were performed in KK mice and DD mice at the age of one week through 16 months. The KK mice, in general, revealed a less increase in body weight, compared with the control mice. Impaired glucose tolerance in the KK mice was observed at the age of 2 weeks and remained for 12 months. The width of MCBM increased in both KK and DD mice with the aging process until 6 months. Thereafter, however, the KK mice revealed a significant increase of the width of MCBM. In both KK mice and DD mice, significant correlations were observed between age and the width of MCBM. In contrast, there was no significant correlation between glucose intolerance figured out as the sum of blood glucose levels and the width of MCBM. The present study suggests that the thickening of MCBM in the KK mice may occur as an aging phenomenon on the one hand and may develop as the consequence of long-term carbohydrate derangement based on genetic disposition on the other hand.

Effect of chondroitin sulfate A in combination therapy with mitomycin C on sarcoma 180 ascites tumor.

Chondroitin sulfate A (CSA) was found to show a strong synergistic effect with mitomycin C in combination therapy of sarcoma 180 ascites tumor implanted in dd mice. Namely, 50% of mice each implanted with 1×106 tumor cells of ascitic form were found to survive on combination treatment with mitomycin C (0.3 mg/kg/day, 5 times) and CSA (1 and 10 mg/kg/day, 5 times). Enhancement of acid phosphatase and β-D-glucuronidase activities of the peritoneal cells in the mice given the combination therapy was evident as compared with those of mice of the control group.

Immunological studies on sporamycin-treated animals.

Sporamycin showed a remarkable tumor regressive activity against sarcoma-180 with a single 5 mg/kg dose of intravenous administration. This antitumor effect on tumor and host animals was examined immunologically. As the results: (1) When sarcoma-180 tumor cells were used as an antigen macrophage migration inhibition reaction by spleen cells derived from the tumor-bearing mice treated with sporamycin was positive at day 7 approximately 14 after the medication and was negative thereafter. (2) The delayed hypersensitivity tested by the foot-pad reaction was positive in tumor-bearing mice treated with sporamycin, and no decrease of foot pad reaction was observed, whereas this reaction decreased remarkably in non-treated tumor-bearing mice. (3) Sarcoma-180 tumor cells were mixed with spleen cells derived from sporamycin-treated mice, and were inoculated into normal dd mice. The growth of tumor cells was inhibited markedly, but no inhibition of tumor growth was observed in case of spleen cells derived from non-treated tumor bearing mice. (4) Combined treatment of sporamycin with PS-K, an immunopotentiator, showed a remarkable synergistic effect.

Neural control of glycogen content and its diurnal rhythm in mouse pineal cell.

In adult male dd mice, possible mechanisms regulating the glycogen content in the pineal cell were investigated by a semiquantitative histochemical method, with particular reference to the role of the sympathetic innervation. Reserpine, superior cervical ganglionectomy (SCGX), or decentralization of the ganglia (DC), as well as continuous light, prevented the nocturnal decrease in the glycogen content, causing a marked increase, and caused a gradual decrease in the size of the pineal cell. In the SCGX or DC group, the glycogen content reached a peak at 2 days and then decreased gradually. The nocturnal decrease was also prevented by propranolol. Noradrenaline caused a marked decrease in the glycogen content. These findings support the hypothesis that the glycogen metabolism and its diurnal rhythm in the pineal cell are regulated by the sympathetic nerve terminals innervating the pineal gland, presumably by the release of noradrenaline. In addition, the nature of the internal mechanism in the organism generating the pineal glycogen rhythm was examined. Light was considered to induce a phase shift in such a mechanism, but reserpine was not.

Effects of tetraacetyl-bis-dehydroascorbic acid, a derivative of ascorbic acid, on Ehrlich cells and HeLa cells (human carcinoma cells).

Derivatives of ascorbic acid were synthesized, and the studies were made on their effects in Ehrlich ascites carcinoma cells, in regard to the inhibition and the prolongation of survival time as well as on the morphological degeneration in HeLa cells. In a model infection study carried out by using tetraacetyl-bis-dehydroascorbic acid in dd mice infected with Ehrlich cells, it was proved that the prolongation of survival time was nearly double in comparison to the control group mice. Also, it was noted that hypertrophy due to abdominal dropsy and body weight were reduced much more than in the control group. From these results, the inhibiting effect of tetraacetyl-bis-dehydroascorbic acid was confirmed. While in the case of DHA and other derivatives, almost no inhibition and prolongation of survival time were observed. As for HeLa cells in a tissue culture, tetraacetyl-bis-DHA, in a dosage of 125-250 mug/ml, demonstrated definitely its morphological degeration. After 125 mug/ml of tetraacetyl-bis-DHA was added to a tissue culture solution of HeLa cells, the cells were washed and recultured. No growth of the cells was observed. Consequently, this substance was confirmed to be anti-HeLa substance with a low toxicity.