Seasonal trade-offs in cell-mediated immunosenescence in ruffs (Philomachus pugnax)

The immune system is an energetically expensive self-maintenance complex that, given the risks of parasitism, cannot be carelessly compromised. Life-history theory posits that trade-offs between fitness components, such as self-maintenance and reproduction, vary between genders and age classes depending on their expected residual lifetime reproductive success, and seasonally as energetic requirements change. Using ruff (Philomachus pugnax), a bird with two genetically distinct male morphs, we demonstrate here a decrease in male immunocompetence during the breeding season, greater variance in immune response among males than females, immunosenescence in both sexes and male morphs, and a seasonal shift in the age range required to detect senescence. Using a phytohaemagglutinin delayed hypersensitivity assay, we assessed cell-mediated immunity (CMI) of males of typical breeding age during the breeding and nonbreeding seasons, and of a larger sample that included females and birds of a greater age range during the non-breeding period. CMI was higher for breeding-aged males in May than in November, but the increase was not related to age or male morph. In November, mean CMI did not differ between the sexes, but the variance was higher for males than for females, and there were no differences in mean or variance between the two male morphs. For both sexes and male morphs, CMI was lower for young birds than for birds of typical breeding ages, and it declined again for older birds. In males, senescence was detected in the non-breeding season only when very old birds were included. These results, generally consistent with expectations from life-history theory, indicate that the immune system can be involved in multifarious trade-offs within a yearly cycle and along an individual's lifetime, and that specific predictions about means and variances in immune response should be considered in future immunoecological research.     

Is cell-mediated immunity related to the evolution of life-history strategies in birds?

According to life-history theory, the development of immune function should be balanced through evolutionary optimization of the allocation of resources to reproduction and through mechanisms that promote survival. We investigated interspecific variability in cell-mediated immune response (CMI), as measured by the phytohaemagglutinin (PHA) assay, in relation to clutch size, longevity and other life-history traits in 50 species of birds. CMI exhibited significant repeatability within species, and PHA responses in chicks were consistently stronger than in adults. Univariate tests showed a variety of significant relationships between the CMI of both chicks and adults with respect to size, development period and lifespan, but not clutch size or prevalence of blood parasites in adults. Multivariate analyses confirmed these patterns but independent variables were too highly correlated to isolate unique influences on CMI. The positive relationship of chick CMI to nestling period is further complicated by a parallel relationship of chick CMI to the age at testing. However, multivariate analysis showed that chick CMI varies uniquely with length of the nestling period. Adult CMI was associated with a strong life-history axis of body size, development rate and longevity. Therefore, adult CMI may be associated with prevention and repair mechanisms related to long lifespan, but it also may be allometrically related to body size through other pathways. Neither chick CMI nor adult CMI was related to clutch size, contradicting previous results linking parasite-related mortality to CMI and the evolution of clutch size (reproductive investment) in birds.     

Transfer of osteosarcoma-specific cell-mediated immunity in hamsters by rabbit dialyzable leukocyte extracts

We have investigated the transfer of specific cell-mediated immunity (CMI) to osteosarcoma-associated antigens (OSAA) to hamsters with dialyzable leukocyte extracts (DLE) from OSAA-immunized rabbits. The transfer of specific CMI was determined by leukocyte adherence inhibition (LAI) assay and skin testing. DLE was prepared from rabbits immunized with OSAA, purified protein derivative (PPD), or fibrosarcoma cell plasma membrane preparation (FSM). Control DLE was prepared from rabbits injected with 0.85% NaCl. Significant leukocyte adherence inhibition was observed with leukocytes from hamsters that had received OSAA-specific, PPD-specific, and FSM-specific rabbit DLE, when OSAA, PPD, and FSM were used as antigens, respectively. Similarly, significant ear swelling after injection of OSAA, PPD, or FSM was observed only in hamsters that had received DLE from rabbits immunized with OSAA, PPD, or FSM, respectively. These results suggest that CMI specific for OSAA, PPD, or FSM can be transferred to normal hamsters by DLE from immunized rabbits.     

Chronic treatment with estradiol restores depressive-like behaviors in female Wistar rats treated neonatally with clomipramine

Neonatal administration of clomipramine (CMI) induces diverse behavioral and neurochemical alterations in adult male rats that resemble major depression disorder. However, the possible behavioral alterations in adult female rats subjected to neonatal treatment with clomipramine are unknown. Therefore, the aim of this study was to explore the effect of neonatal treatment with CMI on adult female rats in relation to locomotion and behavioral despair during the estrus cycle. Also evaluated was the effect of chronic treatment with E2 on these female CMI rats. We found no effects on spontaneous locomotor activity due to neonatal treatment with CMI, or after 21 days of E2 administration. In the FST, neonatal treatment with CMI increased immobility and decreased active swimming and climbing behaviors. Influence of the ovarian cycle was detected only in relation to climbing behavior, as the rats in the MD phase displayed less climbing activity. Chronic E2 administration decreased immobility but increased only swimming in CMI rats. These results suggest that neonatal treatment with CMI induces despair-like behavior in female rats, but that chronic E2 administration generates antidepressant-like behavior by decreasing immobility and increasing swimming, perhaps through interaction with the serotonergic system.     

Mycobacterium phlei as an oral immunomodulator with Newcastle disease vaccine

Experiments were conducted in chickens to understand the effects of oral immunomodulation. Heat inactivated M phlei, a commensal Mycobacterium and a non-specific immunomodulator, was administered orally prior to live Newcastle disease F (ND F) strain vaccination. In experimental birds it lead to an enhanced cell mediated Immune response (CMI) against the vaccine. There was a reduction in the Haemagglutination inhibiting (HI) antibodies. However, it did not affect the protection against a virulent challenge, as the protection percentage was more or less same in vaccinated birds irrespective of the M.phlei administration. M. phlei administration could not enhance the immune response to inactivated ND F vaccine administered orally. The results indicate that M. phlei favours a CMI response to orally administered live ND F vaccine. It may be of potential use in enhancing CMI against vaccines and a cheaper alternative to costlier recombinant cytokines.     

In vivo assessment of tumor-induced nonspecific suppression of contact sensitivity

Summary Normal BALB/c mice were assessed for 2,4-dinitrofluorobenzene (DNFB)-induced contact sensitivity following adoptive transfer of macrophages (Mo). T cells, or their derived products, from normal or tumor-bearing hosts (TBH). Contact sensitivity (CS) was measured by a quantitative radioisotopic ear assay, a total in vivo system based on localization of IP-injected iodinated human serum albumin ([¹²⁵I]HSA) in the DNFB-challenged ear. Adoptive transfer of low or high doses of TBH T cells or their derived supernatants into normal recipients suppresed their responsiveness, while Mo supernatants enhanced it. Moreover, in all cases adoptive transfer of TBH cells or supernatants resulted in a lower CS response than did their normal counterparts. These results further corroborate our previous in vitro data indicating that T cells, or Mo and T cell soluble products, possess immunoregulatory capabilities in vivo.Abbreviations DNFB 2,4-dinitrofluorobenzene - TBH tumor-bearing host - Mo macrophages - ¹²⁵I-HSA iodinated-human serum albumin - CMI cell-mediated immunity - CS contact sensitivity; Ig, immunoglobulin - C complement     

Immunofluorescence in the Study of Marek''s Disease: I. Detection of Antigen in Cell Culture and an Antigenic Comparison of Eight Isolates 1

The indirect fluorescent-antibody (FA) test was applied to the detection of Marek's disease (MD) antigen in cell culture and antibody in the serum of birds. For the detection of antigen, sera were obtained from birds hyperimmunized with the JM strain of MD. MD antigen could be detected in the nucleus and in the cytoplasm of duck and chick embryo fibroblasts and in those of chick kidney cells infected with material known to contain the MD virus. Uninoculated cultures of chicken cells were always free of MD antigen. When chick kidney cells were infected with a stock cellular preparation of MD virus, infected cells could be detected after 24 hr with the FA test. At this time no cytopathological areas were seen by conventional light microscopy. By 7 days after infection, the same number of infected areas were detected by both methods, and the fluorescent areas coincided with the cytopathological areas. This indicates that the fluorescent areas and the areas with cytopathology are caused by the same agent. A straight-line relationship between the dilution of inoculum and the number of fluorescent or morphological foci obtained indicates that one infectious unit produced one fluorescent or morphological focus. In addition, this time sequence study confirmed the cell association of the virus and demonstrated the cell-to-cell spread of infection. Cell cultures inoculated with eight different isolates of MD were tested in all combinations with sera prepared against the same isolates. The antigens were indistinguishable from one another, indicating that either the strains are antigenically identical or there is a common antigen or contaminant in all of them so that they stained equally well. The FA test can detect MD antigen before cytopathological areas develop in cell culture; however, the small size of the area usually examined precludes its use in initial isolations in which only a small number of infectious units are present in the inoculum. MD-infected cells contain a heat-stable antigen similar to that found in herpes simplex-infected cells.     

Isolation and Characterization of an Isolate (HN) of Marek''s Disease Virus with Low Pathogenicity

A cytopathic agent was isolated and characterized as an isolate of Marek's disease herpesvirus (MDHV) with low pathogenicity, and referred to as the HN isolate. This isolate of MDHV did not cause clinical Marek's disease (MD) or death in a highly susceptible line of chickens within 5 weeks after exposure. Gross lesions of limited extent were noted in a few of the inoculated birds. Microscopic nerve lesions in the inoculated and contact-infected birds were invariably minimal, closely resembling C-type MD lesions.     

Isolation and Characterization of an Isolate (HN) of Marek's Disease Virus with Low Pathogenicity

A cytopathic agent was isolated and characterized as an isolate of Marek''s disease herpesvirus (MDHV) with low pathogenicity, and referred to as the HN isolate. This isolate of MDHV did not cause clinical Marek''s disease (MD) or death in a highly susceptible line of chickens within 5 weeks after exposure. Gross lesions of limited extent were noted in a few of the inoculated birds. Microscopic nerve lesions in the inoculated and contact-infected birds were invariably minimal, closely resembling C-type MD lesions.     

Replication of the Reticuloendotheliosis Virus (Strain T) in Chicken Embryo Cell Culture

Investigations were conducted on the in vitro replication of the reticuloendotheliosis (RE) virus (strain T) in specific-pathogen-free chicken embryo fibroblast (CEF) cultures. Active virus production was detected in the tissue culture fluid 24 hr after infection. When injected into chickens, samples taken 42 hr after infection of the cell cultures killed approximately 50% of the birds at a 1:100 dilution. The RE virus titer remained at this level for 5 days before declining. Cell-free virus preparations from tissue cultures rarely resulted in 100% mortality of the assay birds. The level of cell-associated virus was very low. Evidence that the reticuloendotheliosis was not induced by a mycoplasma was indicated by failure to isolate an organism on PPLO Agar (Difco) and failure of kanamycin or amphotericin B to inhibit multiplication of RE virus in vitro. RE virus appeared to be unrelated to members of the avian leukosis and sarcoma complex. It did not induce resistance in CEF cultures to sarcoma viruses of the A or B subgroup of this complex. Similarly, preinfection of cell cultures with leukosis viruses of the A or B subgroup did not inhibit or reduce the replication of RE virus.     

CD4+ and CD8+ T cell priming for contact hypersensitivity occurs independently of CD40-CD154 interactions

The primary effector cells of contact hypersensitivity (CHS) responses to dintrofluorobenzene (DNFB) are IFN-gamma-producing CD8(+) T cells, whereas CD4(+) T cells regulate the magnitude and duration of the response. The requirement for CD40-CD154 engagement during CD8(+) and CD4(+) T cell priming by hapten-presenting Langerhans cells (hpLC) is undefined and was tested in the current study. Similar CHS responses to DNFB were elicited in wild-type and CD154(-/-) animals. DNFB sensitization of CD154(-/-) mice primed IFN-gamma-producing CD8(+) T cells and IL-4-producing CD4(+) T cells. However, anti-CD154 mAb MR1 given during hapten sensitization inhibited hapten-specific CD8(+), but not CD4(+), T cell development and the CHS response to challenge. F(ab')(2) of MR1 failed to inhibit CD8(+) T cell development and the CHS response suggesting that the mechanism of inhibition is distinct from that of CD40-CD154 blockade. Furthermore, anti-CD154 mAb did not inhibit CD8(+) T cell development and CHS responses in mice depleted of CD4(+) T cells or in CD4(-/-) mice. During in vitro proliferation assays, hpLC from mice treated with anti-CD154 mAb during DNFB sensitization were less stimulatory for hapten-primed T cells than hpLC from either control mice or mice depleted of CD4(+) T cells before anti-CD154 mAb administration. These results demonstrate that development of IFN-gamma-producing CD8(+) T cells and the CHS response are not dependent on CD40-CD154 interactions. This study proposes a novel mechanism of anti-CD154 mAb-mediated inhibition of CD8(+) T cell development where anti-CD154 mAb acts indirectly through CD4(+) T cells to impair the ability of hpLC to prime CD8(+) T cells.     

Metabolic consequences of overlapping food restriction and cell‐mediated immune response in a long‐distance migratory shorebird,the little ringed plover Charadrius dubius

Investment in immunity is commonly viewed as an energetically costly activity in birds. Although several studies have focused on the energy cost of mounting an immune response and its concomitant physiological trade‐offs, nothing is known about the metabolic adjustments experienced by immunochallenged birds under resource limitation, or about the basal metabolism cost of mounting cell‐mediated immune (CMI) responses in bird species other than non‐migratory passerines. Here we measured the basal metabolic rate (BMR), inflammatory response, and body mass in ad libitum fed and food‐restricted little ringed plovers Charadrius dubius challenged with phytohemagglutinin (PHA) in order to assess the energy cost, the strength, and the time course of the CMI response in a long‐distance migratory bird in different nutritional states. We found that ad libitum birds injected with PHA significantly increased both mass‐independent BMR and inflammatory response, whereas birds with an induced food restriction‐immune response overlap experienced a mass‐independent BMR downregulation and decreased inflammatory response relative to ad libitum birds. We suggest that both the BMR downregulation and the diminished inflammatory response observed in birds facing such an overlap could be energy‐saving mechanisms to maintain the body mass above a critical level and maximize fitness.     

Effect of prey availability on the abundance of White‐breasted Wood‐Wrens,insectivorous birds of tropical lowland forests

Some understory insectivorous birds manage to persist in tropical forest fragments despite significant habitat loss and forest fragmentation. Their persistence has been related to arthropod biomass. In addition, forest structure has been used as a proxy to estimate prey availability for understory birds and for calculating prey abundance. We used arthropod biomass and forest structural variables (leaf area index [LAI] and aerial leaf litter biomass) to explain the abundance of White‐breasted Wood‐Wrens (Henicorhina leucosticta), tropical understory insectivorous birds, in six forests in the Caribbean lowlands of Costa Rica. To estimate bird abundance, we performed point counts (100‐m radius) in two old‐growth forests, two second‐growth forests, and two selectively logged forests. Arthropod abundance was the best predictor of wood‐wren abundance (wi = 0.75). Wood‐wren abundance increased as the number of arthropods increased, and the estimated range of bird abundance obtained from the model varied from 0.51 (0.28 – 0.93 [95%CI]) to 3.70 (1.68 – 5.20 [95%CI]) within sites. LAI was positively correlated to prey abundance (P = 0.01), and explained part of the variation in wood‐wren abundance. In forests with high LAI, arthropods have more aerial leaf litter as potential habitat so more potential prey are available for wood‐wrens. Forests with a greater abundance of aerial leaf litter arthropods were more likely to sustain higher densities of wood‐wrens in a fragmented tropical landscape.     

鸡MD基因5'侧翼区多态性与鸡生长和体组成性状的相关研究

苹果酸脱氢酶(Malate Dehydrogenase,MD)是一种氧化还原性酶,参与体内多种能量代谢反应.它可以催化苹果酸氧化脱羧生成丙酮酸和CO2,并使NADP+还原成NADPH,NADPH是脂肪酸合成所必需的载体,棕榈酸可以利用生成的NADPH来合成长链脂肪酸,MD的活性与脂肪酸合成效率之间存在密切的相关,MD也参与体内骨骼肌、心肌的能量代谢,并对肌纤维的生长有一定的调节作用.根据鸡MD基因的5侧翼区序列设计一对引物,用直接测序的方法在侧翼区检测多态性位点,在235bp(GenBank登录号U49693)处发现一个SNP位点,此位点是一个限制性内切酶(SphⅠ酶)发生变化的位点.以东北农业大学高低脂双向选择系的第8世代肉鸡和东农F2资源群体为实验材料,用PCR-RFLP的方法进行基因型分析,建立适合的统计模型,进行基因型与生长和体组成性状的相关分析.结果表明在高低脂系第8世代肉鸡中AA基因型个体的腹脂重和腹脂率显著高于BB基因型个体(P＜0.05);BB基因型个体的大胸肌重和大胸肌率显著高于AA基因型个体(P＜0.05).在东农F2资源家系中BB基因型个体的大胸肌重和大胸肌率显著高于AA和AB基因型个体(P＜0.05);AA基因型个体的肝脏重和肝脏率显著高于BB基因型个体(P＜0.05).综上所述,MD基因可能是影响鸡生长和体组成性状的主效基因或与控制生长和体组成性状的主效基因相连锁.     

Sensitization through carcinogen-induced Langerhans cell-deficient skin activates specific long-lived suppressor cells for both cellular and humoral immunity

Application of 2,4-dinitrofluorobenzene (DNFB) to BALB/c mouse skin depleted of epidermal Langerhans cells (LC) by the chemical carcinogen 7,12-dimethylbenz[a]anthracene (DMBA) activated cells which suppress both contact sensitivity and antibody production when transferred into naive host mice. Tolerance was induced by a concentration of DNFB optimal for inducing contact sensitivity in solvent-treated control mice. The cellular and humoral responses of hosts to a second antigen, 2,4,6-trinitrochlorobenzene (TNCB), were unaffected by these suppressor cells, demonstrating specificity for DNFB. Suppressor cells for cellular and humoral immunity could still be demonstrated 6 months following activation, by which time some mice had died, presumably of old age. The dose responses to sensitizer for generation of cells which suppressed contact sensitivity and antibody production differed, indicating that separate populations of suppressor cells probably inhibit these responses. Hence, during cutaneous chemical carcinogenesis, depletion of LC may allow activation of specific long-lived suppressor cells capable of inhibiting cellular or humoral antitumor immune responses.     

基于集合卡尔曼滤波的南雄烟草LAI数据同化研究

叶面积指数(LAI)是表征烟草生长健康状态的重要指标之一,获取准确的LAI数据是监测烟草生长走势的重要步骤。以广东省南雄地区为试验区开展了集合卡尔曼滤波同化方法在烟草LAI的应用研究。通过野外实测得到南雄烟草生长期内的高光谱数据,并计算每个生长期的归一化植被指数(NDVI),依据NDVI值获得LAI测量数据;通过积温数据和实测LAI数据构建了符合南雄地区烟草LAI变化规律的LOGISTIC模型;并以LAI为研究变量,利用集合卡尔曼滤波数据同化技术融合NDVI数据计算得到的LAI和简化LOGSITIC模型拟合得到的LAI这两种不同的数据信息,获取实验区烟草生长期时间序列上的连续LAI数据。最后,进一步对比了数据同化方法、NDVI计算LAI方法和LOGISTIC模型拟合这3种方法获取烟草LAI的效果。结果显示:数据同化方法、NDVI计算LAI方法和LOGISTIC模型拟合3种方法均可一定程度上表征烟草LAI的变化状态,其中数据同化方法拟合效果最优。实验发现NDVI计算LAI方法在烟草生长前后期LAI值出现偏大或偏小的异常情况;LOGISTIC模型拟合则不能有效的描述烟草LAI的突发性变化;同化方法综合作物生长模型和遥感监测的优势,能够动态调节参数得到LAI优化结果,同化后LAI结果和真实值吻合,变化曲线更符合烟草的实际生长状况。     

Domestication effects on foraging strategies in fowl

Birds of two different breeds differing in degree of domestication were studied to reveal any differences in foraging strategies between them. The breeds were wild-type birds (crossing between red jungle fowl (Gallus gallus) and Swedish bantam (Gallus gallus domesticus) and domestic birds (Swedish bantam), breeds representing an increasing level of domestication. Bantam birds have not been selected for any specific characteristics. The birds were allowed to forage in an experimental pen containing two separate food patches, which depleted as a function of being exploited, to see how well the different breeds were able to assess costs and benefits as the distance between patches were changed (short distance between patches compared to long distance between patches). Both breeds behaved in accordance with some general predictions of optimal foraging theory, i.e. moved between patches, left patches before these were empty and stayed for a shorter time in more depleted patches. Wild-type birds responded more than domestic birds to an increase of distance between patches, by spending longer average time in patch when there was a long distance between them compared to when there was a short distance. The wild-type birds adopted what seemed to be a more costly foraging strategy, moving more between patches than the domestic birds without ingesting more feed. During domestication, in the protected environment provided by man, individuals using less costly behavioural strategies may have gained increased fitness over those spending more energy on foraging. Although domestic birds still possessed the ability to respond adaptively to environmental conditions, the differences between the wild-type and the domestic breed might be a result of the reduction of the natural selection pressure which accompanies domestication.     

Inhibition of the T suppressor circuit of delayed-type hypersensitivity by interferon

The effects of electrophoretically pure murine interferon (Mu-IFN-alpha beta) on the T suppressor pathway and on the T effector cell of delayed hypersensitivity (TDH) were investigated in BALB/c mice, in a 2,4-dinitrofluorobenzene (DNFB) contact-sensitivity model. Various T cell subpopulations, suppressor T cells of the afferent (Ts-aff) and efferent (Ts-eff) types, an auxiliary Ts (Ts-aux), as well as TDH were induced, and their function was assessed in transfer experiments. The results were as follows. At a dose of 5 X 10(3) U, IFN was shown to inhibit the Ts-aff response, when given to the donor animal shortly after induction of the Ts-aff subpopulation or when injected into the recipient 2 hr after spleen cell transfer. Pretreatment in vitro with IFN of the splenic cells to be transferred also abolished the Ts-aff response. Similar amounts of IFN were able to inhibit the generation of Ts-eff in the donor animals, whereas 10-fold-higher amounts were needed in vivo or in vitro to block the functional expression of Ts-eff in the recipient animal. Intravenous injection of IFN into recipients of Ts-eff on day 0 and 1 after sensitization inhibited the expression of the Ts-eff transferred 1 day before ear challenge. This suggests that the Ts-aux response required for the TDH suppression by Ts-eff is blocked by IFN. Secretion of a suppressor factor by Ts in vitro was not blocked by IFN. Treatment of the donor of suppressor factor-secreting Ts with IFN, however, blocked the induction of this Ts. The TDH were not sensitive to IFN even at amounts approximately 100 times higher than those used for the Ts inhibition in vivo as well as in vitro. These results demonstrate that low amounts of IFN may selectively block the suppressor pathway, because induction of these regulatory T cell subsets appears to be particularly sensitive to IFN. The exact mechanism of the IFN-mediated inhibition of Ts is not yet clear. The data suggest an important regulatory function of IFN in delayed-type hypersensitivity (DTH) reactions.     

Tolerance and contact sensitivity to DNFB in mice. V. Induction of tolerance with DNP compounds and with free and membrane-associated DNFB.

Immunologic unresponsiveness (tolerance) was induced in a mouse model of contact sensitization to DNFB. The ability to induce tolerance varied with the chemical reactivity of the tolerogen; DNFB was highly tolerogenic, DNBSO3 was moderately tolerogenic, and DNP-lysine was not tolerogenic. Although DNFB is considered a highly reactive compound, tracer studies of injected DNFB showed that it was rapidly excreted. Further studies were therefore done with DNFB attached to mouse erythrocytes. Tolerance to DNFB-RBC was highly specific in vivo; mice tolerant to DNFB showed normal reactivity to TNCB (picryl chloride.) Cells of mice tolerant to DNFB-RBC were also unresponsive to DNBSO3 in vitro. Tolerance to DNFB, DNBSO3, and DNFB-RBC all required time to develop, suggesting that an active process was involved.     

On the activity of trifluoperazine and palmitoylcarnitine in mice: delayed hypersensitivity models

The effect of pre- and post-challenge treatments with trifluoperazine and palmitoylcarnitine, two protein kinase C (PKC) inhibitors characterised by their interaction with the phospholipid enzyme cofactor, on the inflammation caused by delayed hypersensitivity (DTH) to dinitrofluorobenzene (DNFB) and sheep red blood cells (SRBC) in mice is reported. The activity of dexamethasone and two immunosuppressors, azathioprine and methotrexate, is also evaluated. The effectiveness of pre-treatment with each of the test drugs diminished when the DNFB challenge dose increased, whereas trifluoperazine and azathioprine were more active when administered after the challenge at the high DNFB dose. Trifluoperazine, which is also a calmodulin-antagonist, was the more effective of the PKC inhibitors tested on DNFB-DTH (39% and 59% inhibition swelling 24 and 96 h after challenge, respectively). SRBC-DTH was sensitive only to the action of the drugs given after challenge. In this test, PKC inhibitors showed a moderate effect, in the same range as methotrexate, whereas dexamethasone suppressed the reaction. The ability of trifluoperazine in inhibiting cutaneous DTH reaction, depending on the treatment schedule and the hapten challenge dose, has been determined.