Response Of Spleen Lymphocytes Research Articles

Developmental Exposure To Low Doses Of Dichlorodiphenyltrichloroethane Disrupts Functional And Morphologic Maturation Of The Spleen In Prepubertal Rats

Dichlorodiphenyltrichloroethane (DDT) is a versatile persistent pollutant with endocrine disrupting properties and an alleged immune modulator. Objective — to evaluate the parameters of functional and morphological maturation of the spleen in prepubertal rats exposed to low doses of DDT. Material and Methods — From the moment of mating, during pregnancy and lactation, female rats received a solution of o,p-DDT (20 μg/L) instead of tap water. The offspring of intact rat dams were considered the control group. Male DDT-exposed and control rats were sacrificed on postnatal day 7. The spleens were surgically removed under sterile conditions. Anatomical and histological examination of the spleen, ex tempore proliferation of splenocytes, and assessment of splenic T cells and their ability to respond to the mitogen concanavalin A were assessed. Results — Rats exposed to DDT exhibited a significantly reduced proliferative response of spleen lymphocytes to the mitogen concanavalin A. However, morphological evaluation revealed no differences in spleen anatomy between control and exposed rats. Histological examination exposed accelerated development of lymphoid tissue in the spleen of rats exposed to DDT. The ex-tempore proliferation test yielded a higher rate of mitotic division of splenocytes in exposed rats. In contrast to controls, they had a lower percentage of T cells in their spleen. Conclusion — Developmental exposure to low doses of the endocrine disruptor DDT impairs functional and morphological maturation of the spleen in prepubertal rats. DDT accelerates the formation of lymphoid compartments and weakens the functional maturation of the spleen as the organ with an immune function.

Modulatory effects of feeding pregnant and lactating mice Rhodiola kirilowii extracts on the immune system of offspring.

Plants of Rhodiola genus are medicinal herbs that have a number of therapeutic properties, including anti-inflammatory and immunomodulatory activity. The present study aimed to determine whether the use Rhodiola kirilowii as an immunostimulant during pregnancy has an adverse effect on the development of the offspring immune system. Following mating, pregnant mice were placed in three groups that were fed during pregnancy and lactation with R. kirilowii aqueous extract (RKW; 20 mg/kg), R. kirilowii 50% hydro-alcoholic extract (RKW-A; 20 mg/kg) or water (control group), receiving water. Following birth, offspring were given six weeks to develop prior to evaluation of their immune system. Morphometric and morphological examination of the spleen did not reveal any abnormalities or differences between the experimental and control groups. However, both RKW and RKW-A splenic lymphocytes presented a diminished proliferative response to concanavalin A. RKW spleen lymphocytes demonstrated increased metabolic activity following phytohaemagglutinin (PHA) stimulation, which was associated with a higher percentage of cluster of differentiation 4 positive spleen cells and lower interleukin-17a (IL-17a) serum concentration. The RKW-A group exhibited a diminished proliferative response of spleen lymphocytes to PHA and lipopolysaccharide (LPS), and increased serum concentrations of IL-10 and tumor necrosis factor-α (TNF-α). The progeny of mice fed with RKW-A extract demonstrated a significantly lower level of anti-SRBC antibody following immunization compared with progeny of the control (P=0.0305) and RKW (P=0.0331) groups. In conclusion, caution is recommended in the use of RKW and RKW-A extracts as immunostimulants in pregnancy.

Effect ofTrichinella spiralisInfection on the Immune Response to HBV Vaccine in a Mouse Model

Vaccination is the most effective and cost-effective way to treat hepatitis B virus (HBV) infection. Collective data suggest that helminth infections affect immune responses to some vaccines. Therefore, it is important to reveal the effects of helminth infections on the efficacy of protective vaccines in countries with highly prevalent helminth infections. In the present work, effects of Trichinella spiralis infection on the protective efficacy of HBV vaccine in a mouse model were investigated. This study demonstrated that the enteric stage of T. spiralis infection could inhibit the proliferative response of spleen lymphocytes to hepatitis B surface antigen (HBsAg) and lead to lower levels of anti-HBsAg antibodies, interferon-γ, and interleukin (IL)-2, along with higher levels of IL-4 and IL-5. However, these immunological differences are absent in the muscle stage of T. spiralis infection. The results suggest that the muscle stage of T. spiralis infection does not affect the immune response to HBV vaccination, while the enteric-stage infection results in a reduced immune response to HBsAg.

The Roles of Macrophages and Nitric Oxide in Interleukin-3-Enhanced HSV-Sr39tk-Mediated Prodrug Therapy

The herpes simplex virus thymidine kinase/ganciclovir (HSV-sr39tk/GCV) system is a well-established prodrug system used in cancer gene therapy. However, this system is currently not effective enough to eradicate malignant tumors completely. This study aimed to evaluate whether co-expression of interleukin-3 (IL-3) could enhance the anti-tumor activity of HSV-sr39tk/GCV prodrug gene therapy using a murine TRAMP-C1 prostate tumor model. In vitro results demonstrated that HSV-sr39tk-transfected cells exhibited enhanced sensitivity to the GCV prodrug, which was not affected by co-expression of the mIL-3 gene. However, in vivo studies showed that co-expression of the mIL-3 gene significantly increased the HSV-sr39tk/GCV-induced tumor growth delay and even cured the tumor. The TRAMP-C1-specific immune response of spleen lymphocytes from mice bearing HSV-sr39tk- and IL-3-expressing TRAMP-C1 tumors was measured by ELISA. Results showed that IL-3-activated IL-4-dominant lymphocytes became IFN-γ- dominant lymphocytes after combined HSV-sr39tk/GCV therapy. The efficacy of combined therapies on tumor regression was reduced when macrophages populations were depleted by carrageenan or NO production was inhibited by administration of the iNOS inhibitor, L-NAME. These results suggest that utilizing a bicistronic vector to express HSV-sr39tk and the IL-3 gene induced an enhanced macrophage- or NO-dependent anti-tumor effect.

Study on the specific immunity induced by dendritic cell vaccine loading allogenic microvascular endothelial cell bEnd.3 antigen against U14 cervical cancer cell in mice

To explore the specific cellular and humoral immunity induced by dendritic cells (DC) vaccine loading allogenic microvascular endothelial cell bEnd.3 antigen against U14 cervical cancer cell of mice. Mouse brain microvascular endothelial cell bEnd.3 was cultured and identified for preparation endothelial cell bEnd.3 antigen. The level of mRNA expression of vascular endothelial growth factor receptor 2 (VEGF-R₂) and integrin αV was detected by reverse transcription (RT)-PCR. The BALB/c mice were immuned with DC loading bEnd.3 antigen 4 times in 4 weeks (bEnd.3-DC group), while the mice only were immuned with DC or injected with phosphate buffer saline (PBS group) as control group. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, cytotoxic T lymphocyte (CTL) response of spleen lymphocytes in vitro, the percentage of CD₃+CD₈+ surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and western blot. The expression of VEGF-R₂ and integrin αV gene in bEnd.3 cells were expressed highly. After the vaccine was injected, the tumors of mice in PBS group grew faster than those in other groups, while the tumors in bEnd.3-DC group grew slowly and disappeared after 2 weeks. The volume of tumors in DC group grew slower than those in PBS group [(0.11 ± 0.13) cm³ versus (3.38 ± 0.34) cm³]. The CTL response of spleen lymphocytes in vitro showed that bEnd.3-DC cells could kill bEnd.3 cells, the special lysis rate was more than 60%. The percentage of CD₃+CD₈+ spleen lymphocytes in bEnd.3-DC group [(38.6 ± 0.7)%] was higher than those in other groups (P < 0.05). The titer of serum antibody of bEnd.3-DC group was 1:3200, while it was 1:800 in DC group and there were not any in PBS group. Immunocytochemistry analysis indicated there were specific antigen-antibody reaction to bEnd.3 cell in bEnd.3-DC group. Western blot analysis revealed that there were specific bands at 220,000 (VEGF-R₂). bEnd.3-DC vaccine can inhibit the tumor growth of U14 cervical cancer cell of mice, which indicates that the special cellular and humoral immunity are induced by bEnd.3-DC antigen which maybe have some antigens in bEnd.3 cells that reacts with endothelial cell proliferation-related antigens.

Gene Cloning, Expression, and Immunological Characterization of Kexin in RatPneumocystis carinii

Objective: Pneumocystis is an opportunistic pathogen that often causes fatal pneumonia in immunocompromised patients. It remains an important cause of morbidity and mortality in immunocompromised hosts. Therefore, the development of a vaccine is needed. Kexin is a furin-like protease which may be involved in the proteolytic processing of other proteins of Pneumocystis. We focused on the cellular and humoral responses to the kexin antigen fragment of Pneumocystis in our study. Methods: The full-length DNA sequence of the kexin gene was amplified from rat Pneumocystis carinii (P. carinii). The recombinant plasmid pET-kexin was constructed, and the recombinant antigen Hiskexin was purified, respectively. Then the mice were immunized with His-kexin. To ensure the isolated protein used in the next experiments was purified and of the right molecular weight, Western blotting analysis was performed. Then the specific humoral immunity was measured by ELISA. The specific cellular immunity was measured by the proliferation response of spleen lymphocytes. Results: The full-length Pneumocystis kexin gene was cloned successfully. The sera could recognize another recombinant antigen GSTkexin containing the identical amino acid sequence of kexin fragment with His-kexin, as identified by Western blotting and ELISA. The splenocytes collected from the mice immunized with His-kexin had significantly proliferative responses to GST-kexin compared to the control animals. Conclusions: These findings showed that kexin was the potential antigen to induce immune defense against P. carinii infection. The result would provide basis for the further vaccine study in the development of immunoprophylaxis and immunotherapy against P. carinii infection.

Effects of drying and extrusion on colour, chemical composition, antioxidant activities and mitogenic response of spleen lymphocytes of sweet potatoes

Abstract Antioxidative and physiological property changes of freeze-dried, hot air-dried and extruded products made from two different colours of sweet potatoes (yellow and orange) were investigated. The results showed that there were no significant differences of dietary fibre content amongst all the treatments, except that the extruded products of orange sweet potatoes had higher dietary fibre contents. However, the water solubility index (WSI) values and antioxidant content were significantly different between yellow and orange sweet potatoes, and significantly different amongst freeze-dried, hot air-dried and extruded samples. The WSI values of the extruded samples were significantly higher than those of the hot air- and freeze-dried samples. The freeze-dried samples of orange sweet potatoes had more total phenolic compounds, β-carotene, and anthocyanin and had better scavenging effect on DPPH radicals than had freeze-dried yellow sweet potatoes. However, the scavenging effect on DPPH radicals was appreciably greater for hot air-dried than freeze-dried yellow sweet potatoes, but this trend was opposite for orange sweet potatoes. The extrusion process significantly increased the WSI values, scavenging effect on DPPH radicals and total phenolic compounds but decreased the β-carotene and anthocyanin for both yellow and orange sweet potatoes. At lower concentration (25–50 μg/ml), the methanolic extract of freeze-dried yellow sweet potatoes possessed a greater capacity of increasing the mitogenic response than did freeze-dried orange sweet potatoes; at higher concentration (100–200 μg/ml), the proliferation of the lymphocytes increased when stimulated with mitogen Con A for all the samples. This suggests that mitogen Con A selectively promotes T-cell-dependent proliferative activity.

Immunorestoration and augmentation of mitogen lymphocyte response in Graffi tumor bearing hamsters by purified saponin mixture from Astragalus corniculatus

Investigations on the effect of purified saponin mixture (PSM) obtained from the aerial parts of Astragalus corniculatus Bieb. (Fabaceae) on mitogen response of the spleen cells in Graffi tumor bearing (GTBH) and healthy hamsters were reported. The saponin mixture in a doses of 50mg/kg b.w. was injected i.p. 4 times starting simultaneously with implantation of tumor cells. Stimulation indices to phytohemagglutinine (PHA) and lipopolysaccharide (LPS) of lymphocytes in spleens of tumor bearing hamsters (TBH) were significantly decreased during the whole period of the observation. It was established that PSM stimulated the functions of spleen cells in Graffi-TBH, resulting in increased mitogen response to PHA and LPS. The stimulation was better expressed in healthy PSM-treated hamsters. The proliferation response of spleen lymphocytes to PSM was also found. PSM did not change the in vitro proliferation ability of Graffi tumor cells. The results obtained proved the immunostimulating and immunorestorating activity of PSM on the T- and B-spleen cells in healthy and GTBH hamsters, as well as the proliferative response of it to PSM.

Effect of Dietary Restriction on DNA Synthesis in Vitamin E‐Deficient Rats

To assess the effect of dietary restriction on increased oxidative stress conditions, we measured the proliferative response of spleen lymphocytes from the following groups of adult rats: (1) control fed ad libitum (14 months of age); (2) vitamin E-deficient (12 months of age); and (3) vitamin E-deficient maintained on dietary restricted paradigm, that is, every other day schedule (12 months of age) animals. No significant change was observed among the three groups investigated at 24 h. At 48 h, [(3)H]thymidine incorporation was significantly lower in vitamin E-deficient rats vs. the other groups at Con A concentrations of 1 and 5 mug/mL, while at Con A concentration of 10 mug/mL the incorporation of the labeled compound in lymphocytes was significantly lower than only the vitamin E-deficient rats vs. controls. At 72 h: nonstimulated lymphocytes from ad libitum fed control rats showed significant higher values of [(3)H]thymidine incorporation vs. the other groups; no significant difference was found among the three groups investigated at 1 and 10 mug/mL Con A concentrations, while at 5 mug/mL Con A concentration, the lymphocytes from vitamin E-deficient rats showed a significant lower value of [(3)H]thymidine incorporation vs. the other groups. These data support that vitamin E-deficiency impairs the proliferative response of spleen lymphocytes from adult rats, while dietary restriction appears to be able to reverse this alteration. Although the mechanism(s) of action of dietary restriction in prolonging the life span and ameliorating health conditions are not know, it is currently supported that a reduced food intake results in a better control of free radical attacks to biological molecules as well as to several cellular and system functions. With specific reference to the present findings, dietary restriction may help the mitotic process dynamics to be accomplished in a condition of low rate of free radical damage, thus representing a physiological intervention capable of modulating positively the proliferative capacity of spleen lymphocytes and, in turn, the immune system, even in adverse conditions such as increased oxidative stress.

Effect of cyclosporin A in Lewis rats in vivo and HeLa cells in vitro.

The aim of this study was to compare the effect of cyclosporin A (CsA) in inbred Lewis rats with published assessment of immunotoxicity in 'classical' outbred Wistar rats. A second purpose was to consider the contribution of a panel of in vitro assays in cell cultures when added to an immunotoxicity study in vivo. The in vivo effect of CsA was investigated in a 28-day subacute immunotoxicity study in male Lewis rats at three different concentrations: 1.25, 5 and 20 mg kg(-1). The highest dose of CsA exceeded the maximum tolerated dose. A drop in body, spleen and popliteal lymph node weight of exposed animals displayed symptoms of toxicity. At a high toxic dose, haematological changes showed a decrease in the leucocyte count and in the percentage of lymphocytes, and an increase in the percentage of polymorphonuclear leucocytes. The haematocrit was significantly dose-dependently suppressed in all rats exposed to CsA. A similar dose-dependent depression of the mean cell volume of erythrocytes was found in rats given high and middle doses of CsA. The phagocytic activity of polymorphonuclear leucocytes and monocytes also was significantly dose-dependently suppressed. No significant changes in primary antibody response to sheep erythrocytes or in vitro proliferative response of spleen lymphocytes to mitogens were found in those rats.A battery of in vitro cytotoxicity methods was selected for the evaluation of metabolic and functional activity of subcellular organelles (mitochondria, lysosomes) and for the detection of drug-induced superoxide-mediated damage in HeLa cells. This cell line was chosen because it has a lower activity of superoxide dismutase (SOD) than normal cells and is sufficiently sensitive for the detection of the induction of oxygen radicals. The in vitro results indicated a direct relationship between CsA cytotoxicity and a change in the mitochondrial enzyme activity, as well as an induction of superoxide production. The results of the study indicated that a combination of selected in vivo and in vitro methods is an inexpensive way to obtain more complex information on cell status affected by xenobiotics.

Adaptive Immune Responses of Legumin Nanoparticles

Legumin is one of the main storage proteins in the pea seeds (Pisum sativum L.) and the molecules of this protein have the capacity of binding together to form nanoparticles after aggregation and chemical cross-linkage with glutaraldehyde. The aim of this work was to study the adaptive immune response of legumin nanoparticles in rats. Following intradermal immunisation with the native protein legumin and legumin nanoparticles of about 250 nm, the humoral and cell-mediated immune responses were analysed in rats. The humoral responses against legumin and legumin nanoparticles were examined by western blot and ELISA analysis. Both techniques clearly showed that sera from rats immunised with legumin strongly expressed antibodies against this protein. On the contrary, serum samples from rats inoculated with legumin nanoparticles did not contain detectable amounts of antibodies. These results may be explained by a reduction on the antigenic epitopes of the protein induced by the glutaraldehyde used during the cross-linking step. Concerning the cell-mediated response, neither legumin nor legumin nanoparticles stimulated an immunogenic response. This absence of response of spleen lymphocytes for legumin and legumin nanoparticles may be explained by a cytostatic effect of legumin which was corroborated by the evaluation of the middle phase of cell apoptose. In fact, both legumin and legumin nanoparticles are potent inductors of a cytostatic phenomenon and showed a significant increase of the chromatin condensation (p <0.05) as compared with control.

Recombinant Mycobacterium bovis BCG producing the circumsporozoite protein of Plasmodium falciparum FCC-1/HN strain induces strong immune responses in BALB/c mice

The current vaccine against tuberculosis, Mycobacterium bovis strain bacillue Calmette–Guerin (BCG), offers potential advantages as a live, innately immunogenic vaccine vehicle for expression and delivery of protective recombinant antigens. Malaria is one of the severest parasitic diseases in humans especially in the developing world. No efficacious vaccine is currently available. However, circumsporozoite protein (CSP) is a malaria vaccine candidate currently undergoing clinical trials. We analyzed the immune response to recombinant BCG (rBCG) vaccine expressing Plasmodium falciparum CSP (BCG-CSP) under the control of heat shock protein 70 promoter in BALB/c mice. The lymphocytes proliferative response to P. falciparum soluble antigen was significantly higher than those in the groups of BCG and normal saline, and the production of cytokines (IFN-γ and IL-2) in response to malaria antigen was significantly higher in rBCG and BCG groups than control group of normal saline. A specific IgG antibody response against P. falciparum antigen of CSP was also characterized. The booster injection could enhance the production of cytokine, proliferation responses of spleen lymphocytes and the antibodies titer of BCG-CSP. The results in the study demonstrated that rBCG vaccine producing CSP is an appropriate vaccine for further evaluation in non-human primates.

Immunotoxicity of ethyl-4-isothiocyanatobutanoate in male Wistar rats

The immunotoxicity of ethyl-4-isothiocyanatobutanoate (E-4IB) using different immuno-pathological parameters and immune function assays in male Wistar rats was evaluated. The rats were administered intraperitoneally 12 times with E-4IB in three varying doses of 21, 28 and 35 mg/kg of body weight, over a period of 36 days. The doses of E-4IB were set according to the results of previous experiments by its anti-proliferative activity in vivo. High and medium doses of E-4IB exceeded the maximum tolerated dose after the 36-day treatment period. Symptoms of toxicity were displayed by a drop in body weight, spleen and thymus weight and in organ and bone marrow cellularity. Haematological changes displayed a dose-dependent decrease in the percentage of lymphocytes and dose-dependent increase in the percentage of polymorphonuclear leukocytes in peripheral blood. The white blood cell count in rats exposed to a high dose of E-4IB was suppressed. The immune system of rats administered 21 mg/kg of E-4IB (low dose) was unaffected. No changes in primary antibody response to sheep erythrocytes, in vitro proliferative response of spleen lymphocytes to mitogens and phagocytic activity of leukocytes were found in those rats. Our findings indicate that this newly developed anti-cancer drug is not immunotoxic.

Immunomodulatory effects in Litomosoides sigmodontis -infected Mastomys coucha

The levels of parasite-specific IgG1 and IgG2 antibodies and mitogen-induced and parasite-specific proliferative T-cell responses were determined in Litomosoides sigmodontis-infected Mastomys coucha throughout an observation period of 400 days post infection (p.i.). These were compared with the respective reactions in animals that had been immunized with intrauterine stages/microfilariae of the parasite and in animals that had been challenged after immunization as determined at up to 60 days after challenge. IgG1 antibodies to adult antigen developed early and reached a plateau at 120 days p.i., whereas IgG2 antibodies were not found before day 60 p.i., increased with rising parasitemia, reached a plateau at 200 days p.i., and, in some animals, even became the predominant IgG subclass. Proliferative responses of spleen lymphocytes to concanavalin A (Con A) and lipopolysaccharides (LPS), but not Con-A-induced interleukin 2 (IL-2) production, were found to be suppressed in infected animals during patency as compared with uninfected controls. Spleen cells of infected animals showed a weak proliferative reaction to male antigen but were unresponsive to female and microfilarial antigen during prepatency and early patency. Subsequently, when microfilaremia decreased (200 days p.i.), continuously increasing responses to all antigens were observed. Immunized M. coucha developed specific IgG1 and IgG2 antibodies, and their spleen cells showed strong proliferative responses to the three L. sigmodontis antigens. Challenge infections down-regulated the proliferative responses of spleen cells to filarial antigens as early as during the prepatent phase of the challenge infection but supported existing IgG1 and IgG2 responses.

Long-term melatonin supplementation does not recover the impairment of natural killer cell activity and lymphocyte proliferation in aging mice

In this study we evaluated the effect of long-term melatonin (MEL) treatment on the cytotoxic activity and number of natural killer (NK) cells and the proliferative response of spleen lymphocytes to phytohemagglutinin (PHA) or interleukin-2 (IL-2) in old mice. Seventeen-eighteen month-old Balb/c mice were supplemented with MEL (40–50 μg/day/mouse) and sacrificed after eight months. The MEL supplementation was unable to recover the low levels of both endogenous and IL-2-induced NK cell activity found in old untreated mice Also the NK cell number was unaffected by MEL treatment. The spleen lymphocyte proliferative response to both PHA and IL-2 was not different in old MEL-treated compared to old untreated mice. These results indicate that long-term MEL supplementation does not recover the age-related deterioration of NK cell activity and lymphocyte proliferative capacity.

Trypanosoma cruzi: The major cysteinyl proteinase (cruzipain) is a relevant immunogen of parasite acidic antigens (FIII)

This study examined the immune responses induced by cruzipain, a well-characterized T. cruzi antigen, to determine whether it is a relevant immunogen among the parasite acidic antigens (FIII), for which some biological properties have been studied previously. Humoral and cellular immune responses were investigated in BALB C mice after immunization with cruzipain or FIII. Skin tests revealed immediate type-hypersensitivity (ITH) and delayed-type hypersensitivity (DTH) reactions to cruzipain in both groups of immunized mice. IgG1 and IgE isotypes against cruzipain were detected by ELISA in both groups and immunoblot studies showed that these antibodies recognized a major protein band of 50 kDa, cruzipain. The antigen-specific proliferative responses of spleen lymphocytes from both groups of immunized mice were also increased. Immunization with cruzipain or FIII antigen significantly enhanced the percentage survival of mice challenged with 10 3 trypomastigotes. The results revealed high cross-reactivity between cruzipain and FIII, suggesting that cruzipain is a relevant immunogen among the parasite acidic antigens.

Immunosuppression and feeding success of Ixodes ricinus nymphs on BALB/c mice

The effect of repeated infestations of Ixodes ricinus (L.) nymphs on BALB/c mice was studied. Four successive infectations resulted in an increase of tick feeding success. Tick yield and mean engorged weight increased and the length of the feeding period was reduced significantly (P < 0.05-0.01). The increase of specific anti-tick antibodies was not significant (P > 0.05). The blastogenic response of spleen lymphocytes to T-cell mitogens (Con A and PHA-P) was unimpaired or slightly enhanced, whereas the response to B-cell activators (LPS and PWM) was suppressed, as was the total antibody generation in vitro. The numbers of mast cells in murine skin at the tick attachment sites slightly decreased during the third infestation. The suppression of B-cell competence and of antibody generation, together with decrease of skin mast cell numbers in tick attachment sits, are considered to be responsible for enhancement of tick feeding success.

Estrogen effects on lymphoid tissue in neonatal and adult female mice.

Outbred female mice of the NMRI strain were treated with daily doses of diethylstilbestrol (DES; 0.1-10 micrograms), estradiol-17 beta (E2; 5 micrograms), testosterone (T; 5 or 25 micrograms), progesterone (P; 100 micrograms), corticosterone (CC; 10 or 50 micrograms), or olive oil (controls) for the first 5 days after birth. Animals were killed on day 6 after birth, at 2, 4, and 8 weeks or at 4 and 6 months. Only CC resulted in reduced thymus index weight (IW, mg organ weight/g body weight) on day 6 while the estrogen-induced reduced IW was secondary to a reduced body weight. Four-week-old and older estrogen-treated females had increased thymus IW (1.8-fold at 8 weeks after 5 micrograms DES/day neonatally) which was still seen at 6 months. Estrogen-induced thymus enlargement also occurred in 8-week-old inbred NMRI females and in females belonging to the BALB/c strain. Thymus IW in 8-week old T-, P- and CC-treated females was as in controls. The leukocyte counts in blood and bone marrow were lower in DES females than in controls. The responses (thymus IW, peripheral blood leukocyte count) of 8-week-old females to ovariectomy and challenge with DES were different in DES females and in control females. The mitogen response of spleen lymphocytes and thymocytes was similar in controls and DES females. An anomaly in the hypothalamic-pituitary-gonad-thymus axis of adult, neonatally estrogen-treated females might explain the thymus enlargement.

Effects of footshock stress upon spleen and peripheral blood lymphocyte mitogenic responses in rats with lesions of the paraventricular nuclei

To assess the role of the hypothalamic paraventricular nucleus (PVN) in mediating stressor-induced immune alterations, male Lewis rats were subjected to a 1-h session of intermittent footshock stress or home cage conditions 6 days after receiving bilateral or sham PVN lesions. Splenic and peripheral blood lymphocyte proliferative responses to the non-specific mitogens, concanavalin A (ConA) and phytohemagglutinin (PHA), were subsequently measured as were plasma corticosterone levels. In sham-operated rats, footshock markedly elevated plasma corticosterone levels and concurrently suppressed the proliferative responses of peripheral blood and splenic lymphocytes. In PVN-lesioned rats, however, the shock-induced suppression of lymphocyte proliferation in the peripheral blood and the elevation of plasma corticosterone were significantly attenuated, while lymphocyte proliferation in the spleen was suppressed below the level of the sham-treated animals. Thus, by utilizing ablation studies, we have determined that the PVN may play a direct role in the alteration of lymphocyte function during stress, and an intact PVN buffers the effect of stress on the responsiveness of spleen lymphocytes to non-specific mitogens.

Humoral and cellular immune response of BALB/c mice to repeated infestations with Ixodes ricinus nymphs

The capacity of female BALB/c mice to mount an immune response and effective resistance to repeated infestations with I. ricinus nymphs was studied. An anamnestic antibody response and transient in vitro responsiveness of spleen lymphocytes to tick antigens were demonstrated in repeatedly infested mice. On the other hand, the response to concanavalin A—a T-cell mitogen, was suppressed at the same time. In the presented experiment, BALB/c mice did not manifest tick resistance after three successive infestations (with a reinfestation period of 2 weeks). The possibility of an infestation-dependent modulation of immune response in BALB/c mice is discussed.