Rat Lymphoid Cells Research Articles

Cotton rat (Sigmodon hispidus) signaling lymphocyte activation molecule (CD150) is an entry receptor for measles virus.

Cotton rats (Sigmodon hispidus) replicate measles virus (MV) after intranasal infection in the respiratory tract and lymphoid tissue. We have cloned the cotton rat signaling lymphocytic activation molecule (CD150, SLAM) in order to investigate its role as a potential receptor for MV. Cotton rat CD150 displays 58% and 78% amino acid homology with human and mouse CD150, respectively. By staining with a newly generated cotton rat CD150 specific monoclonal antibody expression of CD150 was confirmed in cotton rat lymphoid cells and in tissues with a pattern of expression similar to mouse and humans. Previously, binding of MV hemagglutinin has been shown to be dependent on amino acids 60, 61 and 63 in the V region of CD150. The human molecule contains isoleucine, histidine and valine at these positions and binds to MV-H whereas the mouse molecule contains valine, arginine and leucine and does not function as a receptor for MV. In the cotton rat molecule, amino acids 61 and 63 are identical with the mouse molecule and amino acid 60 with the human molecule. After transfection with cotton rat CD150 HEK 293 T cells became susceptible to infection with single cycle VSV pseudotype virus expressing wild type MV glycoproteins and with a MV wildtype virus. After infection, cells expressing cotton rat CD150 replicated virus to lower levels than cells expressing the human molecule and formed smaller plaques. These data might explain why the cotton rat is a semipermissive model for measles virus infection.

Активність інтерферон-залежної 2' ,5' -олігоаденілат-синтетази в лімфоїдних клітинах щурів в умовах трансформованого середовища

Активність інтерферон-залежної 2' ,5' -олігоаденілат-синтетази в лімфоїдних клітинах щурів в умовах трансформованого середовища

Alterations induced in vitro by ochratoxin a in rat lymphoid cells

Ochratoxin A (OTA) is a nephrotoxic mycotoxin produced by species of the genus Penicillium and Aspergillus that is present in food and feed as a natural contaminant. It modifies the immune function in animals and inhibits the proliferative response of lymphocytes in vitro. The toxic effect of OTA (0.5, 2, 20 microM) in lympho-proliferative response, natural killer (NK) cell activity, cytotoxic T lymphocytes (CTL) activity and macrophages' bacteriolytic capability was studied in vitro after 1 hour of treatment. The proliferative response of lymphocytes to concanavalin A and lipopolysaccharide was not affected by OTA; the cytotoxic activity of NK cells was dose-dependent decreased; the CTL activity was significantly decreased at the lowest concentration; the bacteriolytic activity of macrophages varied only slightly. These in vitro results reproduced, at least in part, some effects detected previously in vivo. The protein synthesis inhibition and the oxidative metabolism of OTA coupled to the prostaglandin synthesis are probably implicated in NK cells' toxicity, because the effects were reverted by the addition of phenylalanine or piroxicam to the culture medium. The induction of apoptosis seems to be the principal mechanism of action in the CTL effect. The intracellular concentration of OTA after 1 hour was analysed by HPLC and was found to be proportional to the quantity of OTA added to the culture medium for the three cell types; the presence of phenylalanine and piroxicam on the culture medium did not change the intracellular OTA concentration.

Development of rat CD45+ 13-day-old fetal liver cells in SCID mouse fetal thymic organ cultures.

A phenotypic analysis of the lympho-hemopoietic cells which occur in the liver of 13-day-old fetal rats was achieved by flow cytometry in an attempt to further characterize the rat lymphoid progenitor cells. A small fraction of rat 13-day-old fetal liver (r13FL) cells, which weakly expressed the leukocyte common antigen CD45, constituted a homogeneous Thy-1(hi), CD71(-), CD44(+), MHC class I+, CD43(+) cell subpopulation negative for CD45RC, CD3, TCRalphabeta, TCRgammadelta, CD2, CD5, CD4, CD8, CD25, CD28, NKR-P1a and sIg. On the contrary, the CD45(-) cells were a heterogeneous cell subset which expressed Thy-1, CD71 and CD44 at distinct levels. After MACS separation, the CD45(+) r13FL cells, but not the CD45(-) cell subset, in vitro repopulated 14-day-old SCID mouse fetal thymic lobes providing rat T cells, both TCRalphabeta and TCRgammadelta, NK cells, and thymic dendritic cells but not B lymphocytes. Interestingly, NKR-P1a(lo) TCRalphabeta+ or TCRgammadelta+ cells developed in the xenogeneic cultures, and a rare CD4(+)CD8(+) double-positive subpopulation among the TCRgammadelta-expressing cells accumulated in the oldest cultures. These results are discussed from the double perspective of the nature of the precursor cells which colonize the fetal thymus and the relevance of the xenogeneic SCID mouse fetal thymic microenvironment for supporting rat lymphopoiesis.

IN VIVO CHARACTERIZATION OF CELLULAR XENOIMMUNITY

47 Introduction: Rejection of xenogeneic tissue is mediated by a unique collaboration of both humoral and cellular arms of the immune system. The humoral response has been the topic of considerable investigation while the cellular response remains largely unstudied. In the present work, we characterized the cellular xenogeneic response by following lymphocytes labeled with carboxyflourescein (CFSE), a fluorescent dye that covalently binds intracellular proteins. Because CFSE fluorescence intensity is halved with each cell division, calculation of the rate, number of divisions, and precursor frequency of responding cells is possible. Methods: CFSE labeled splenocytes or lymph node cells were utilized in vivo and in vitro to compare allogeneic and xenogeneic responses. In the in vitro experiments, rat lymphoid cells were stimulated with irradiated xenogeneic cells (Lewis anti-B6). Control responders were stimulated by either Con A or allogeneic mouse cells (B6 anti-BALB/c). For in vivo experiments, Lewis responders were injected into supra-lethally irradiated (2500R) xenogeneic (B6) recipients. Responding cells were collected at designated time points, stained for T cell subset markers (CD4 and CD8) and analyzed by flow cytometry. Results: Rat cells stimulated with Con A had not divided by 48 hrs, but evidenced vigorous proliferation by 72 hrs. The proliferation kinetics showed sequential increases (to a maximum of six divisions), and the calculated precursor frequency of CD4 T cells was constant at all intervals (approx. 70%). At 72 hours, the in vitro xenoresponse yielded a precursor frequency of 0.7% with six cell divisions as compared to 4% and six divisions in allogeneic mouse controls. For in vivo experiments, the xenogeneic precursor frequency was 1.8% with six divisions at 60 hours versus 15% precursor frequency and six divisions allogeneic controls. (Figure)FigureConclusions: The use of CFSE labeling provides a unique tool to monitor the responses of xeno-reactive cells in vivo. We found that compared with allogeneic responses, xenogeneic responses have reduced division kinetics and precursor frequency. The relatively greater estimate of precursor frequencies in the in vivo experiments may result from a relative enrichment of reactive cells in the recipient spleen.

Differential expression of elongation factor-2, α4 phosphoprotein and Cdc5-like protein in prolactin-dependent/independent rat lymphoid cells

The differential display of mRNAs technique was used to identify genes which are differentially expressed in the prolactin (PRL)-dependent Nb2-11C and PRL-independent Nb2-Sp rat lymphoma cell lines. The technique was validated by the differential display of the proto-oncogene c- myc in PRL-treated, but not in untreated, Nb2-11C cells. Nine DNA bands, isolated from the 6% display gels and confirmed by reverse Northerns to be differentially expressed, were cloned and gave rise to ten unique clones. DNA sequencing showed that these clones had limited homologies to known genes or uncharacterized expressed sequence tags. Using one of these clones (6ac1.12) as a probe in Northern analysis, a transcript of approximately 4 kb was detected which was elevated in PRL-treated Nb2-11C cells and in mid-log phase growing Nb2-Sp cells. Similar to c- myc, expression of the 4 kb transcript increased in Nb2-11C cells given PRL for 3 h (±the phorbol ester TPA) but not in cells given TPA alone. The 4 kb transcript also increased with increasing Nb2-11C cell densities. By screening an Nb2-Sp cDNA library with 6ac1.12 as probe, three unique genes were isolated and identified as elongation factor 2 (EF-2), α4 phosphoprotein and a Cdc5-like protein. Each of the three genes were PRL responsive in Nb2-11C cells and expressed constitutively in Nb2-Sp cells. The expression of EF-2 or α4, but not the Cdc-like protein, was dependent on cell densities. EF-2 regulates protein synthesis while the α4 and Cdc5-like phosphoproteins have been implicated in IgG receptor-mediated and mitogen-activated signaling, respectively. The identification that these genes are PRL-responsive and/or differentially expressed in the Nb2-11C and Nb2-Sp cell lines may permit insights into the molecular changes that are involved in regulating the transition to growth factor independence in lymphoid tumors.

The effect of prolactin on casein kinase II, MAP kinase and PKC in rabbit mammary cells and Nb 2 rat lymphoid cells

Prolactin induces milk protein gene expression in rabbit primary mammary cells without any concomitant cell multiplication. Prolactin or other lactogenic hormones is the major inducer of cell division in the rat lymphoid Nb 2 cells. In Nb 2 cells, prolactin also rapidly induces the expression of the c-myc gene, and β-actin and stathmin gene expression is induced more slowly. The possible involvement of casein kinase II (CKII), mitogen-activated protein kinase (MAPK) and protein kinase C (PKC) in these process is not well known. The present work was undertaken to evaluate the effect of prolactin on these protein kinases and to determine the possible involvement of these enzymes in the activity of several genes under the control of the hormone. In rabbit mammary cells, prolactin did not alter CKII activity but did transiently stimulate MAP kinase activity. Prolactin also stimulated Ca 2+-independent PKC. This effect was visible after 10 min and was maintained for at least 24 hr. Staurosporine, an inhibitor of PKC and of several tyrosine kinases altered Ca 2+-independent PKC only moderately. In contrast, GF 109203X, a potent and specific inhibitor of PKC, abrogated almost all PKC activity. Staurosporine, but not GF 109203X, prevented the induction of the casein gene by prolactin. In Nb 2 cells, prolactin induced a slow stimulation of CKII activity. The hormone did not induce MAP kinase activity. Prolactin stimulated Ca 2+-independent PKC over periods of 24 hr. GF 109203X, but not staurosporine, inhibited PKC activity, whereas staurosporine but not GF 109203X, inhibited the induction of Nb 2 cell multiplication and the accumulation of c-myc, β-actin and stathmin mRNAs. From these data, it can be concluded that (1) the stimulation of CKII by prolactin in Nb 2 cells is concomitant with cell multiplication: (2) MAPK stimulation is not necessary for prolactin to induce Nb 2 cell multiplication; and (3) PKC is stimulated in mammary and Nb 2 cells, but this stimulation is not required for prolactin to stimulate casein, c-myc, β-actin and stathmin gene expression and Nb 2 cell division.

Molecular characterization of immune derived proenkephalin mRNA and the involvement of the adrenergic system in its expression in rat lymphoid cells

Proenkephalin (PENK), a classically defined opioid gene, was originally thought to be expressed almost exclusively in the mature nervous and neuroendocrine systems. In the last few years, it was demonstrated, however, that significant levels of PENK mRNA and PENK-derived peptides are transiently expressed in cells of the immune system. Very little is known about the molecular mechanisms regulating this transient expression. In order to investigate those mechanisms, we examined the in vivo expression of PENK mRNA in mesenteric lymph nodes after exposing rats to lipopolysaccharide. In the present study we demonstrate that: (1) promoter usage and splicing of PENK mRNA function similarly in mesenteric lymph nodes as in neural cells; (2) PENK expression in mesenteric lymph nodes is modulated by adrenaline via adrenergic receptors; and (3) the adrenergic system participates in the modulation of the LPS induced PENK mRNA expression. These results provide more evidence for the involvement of opioids in neuro-immune interactions.

Cellular nuclear binding and retention of glucocorticoids in rat lymphoid cells: effect of long-term adrenalectomy.

In the present study we have shown for the first time that lymphocytes from the spleen, thymus and mesenteric lymph nodes can specifically bind and retain 3H-dexamethasone in the nuclear fraction. The use of the specific corticosteroid receptor antagonists, RU28362 and RU38486, for the displacement of labeled dexamethasone revealed that binding was mainly of type-II glucocorticoids receptors. In lymphocytes obtained from mesenteric lymph nodes, binding was significantly higher than in cells obtained from spleen and thymus. Long-term adrenalectomy (up to 30 days), which markedly elevated serum ACTH, caused a time-dependent increase in binding in all lymphoid cells. No increase in binding was found in lymphocytes obtained from rats hypophysectomized for 15 days, in which neither serum ACTH nor corticosterone could be measured. These results suggest that the cellular nuclear binding in lymphocytes may be regulated by a pituitary factor.

Stathmin gene expression in mammary gland and in Nb 2 cells

Mammary gland growth occurs essentially during pregnancy and induction of milk synthesis is triggered at parturition. Prolactin is mammogenic in vivo but only marginally in vitro. Prolactin induces milk synthesis in vivo and in cultured mammary cells. Prolactin is also strictly required for the multiplication of the rat lymphoid Nb2 cells. Stathmin is an ubiquitous and highly conserved phosphoprotein which seems to be involved in the intracellular mechanisms which trigger cell multiplication and differentiation. In the present study, the concentration of stathmin mRNA has been evaluated during the pregnancy-lactation-weaning cycle in mouse and rabbit. Stathmin mRNA appeared at its highest level during pregnancy and it was almost undetectable during lactation. Prolactin injected into mid-pregnant rabbits induced milk synthesis and this effect was not accompanied by any modification of stathmin mRNA concentration. In cultured primary rabbit mammary cells, prolactin induced casein gene expression without any alteration of stathmin mRNA concentration. In Nb2 cells, prolactin induced a progressive increase of stathmin mRNA concentration. This effect was not significant until after 4 h of prolactin action. These data suggest that stathmin is involved in mammary and Nb2 cell multiplication but may not be necessary for mammary cell differentiation.

Study on the lipid composition of aging fisher‐344 rat lymphoid cells: Effect of long‐term calorie restriction

Study on the lipid composition of aging fisher‐344 rat lymphoid cells: Effect of long‐term calorie restriction

Different types of in vivo induced cytolytic T lymphocytes are all LFA-Ihigh and lack CD45 exon 4 (CD45RA) but show distinct CD45RC profiles.

Due to alternate mRNA splicing of exons 4, 5 and 6 (or A, B and C respectively), the CD45 cell surface glycoprotein is structurally heterogeneic in lymphoid cells of different lineage or stage of activation. Previous studies show that in vivo induced allo- and superantigen reactive rat cytolytic T lymphocytes (CTL) preferably belong to the CD45RClow subset, whereas tumour selective CTL express high amounts of CD45RC cell surface molecules. In this paper, reverse transcription polymerase chain reaction technique (RT-PCR) was utilized to evaluate CD45 isoform expression of rat lymphoid cells and in vivo activated rat CTL with distinct specificity and CD45RC profile. Cells from lymphoid organs expressed six CD45 mRNA isoforms, exon(45678), exon(5678), exon (578), exon(678), exon(78) and a novel extensively spliced exon(8) variant. In vivo activated TCR alpha beta + CD8+ cells sorted as CD45RClow expressed exon(78), exon(578) and exon(8), whereas TCR alpha beta + CD8+CD45RChigh cells expressed exon(78), exon(578), exon(5678) and full-length exon(45678). Triple-colour staining indicated high expression of LFA-1 in the cytotoxic CD45RCintermediate and CD45RClow cells, and low expression of LFA-1 in CD45RChigh non-cytotoxic cells from allo- and superantigen activated rats. In contrast, tumour activated TCR alpha beta +CD45RChigh cells were divided in LFA-1high and LFA-1low subsets, and sorting of these subsets revealed that tumour-selective cytotoxicity was confined to the LFA-1high effector cell subset. Furthermore, it was evident that the LFA-1high effector cell subset expressed high levels of exon(5678), exon(578) and exon(78) isoforms and, in contrast to the LFA-1low subpopulation, lacked expression of exon 4 containing full-length CD45 mRNA transcript.(ABSTRACT TRUNCATED AT 250 WORDS)

Induction of donor-specific transplantation tolerance to skin and cardiac allografts using mixed chimerism in (A + B-->A) in rats.

Mixed allogeneic chimerism (A + B-->A) was induced in rats by reconstitution of lethally irradiated LEW recipients with a mixture of T-cell depleted (TCD) syngeneic and TCD allogeneic ACI bone marrow. Thirty-seven percent of animals repopulated as stable mixed lymphopoietic chimeras, while the remainder had no detectable allogeneic chimerism. When evaluated for evidence of donor-specific transplantation tolerance, only those recipients with detectable allogeneic lymphoid chimerism exhibited acceptance of donor-specific skin and cardiac allografts. Despite transplantation over a major histocompatibility complex (MHC)- and minor-disparate barrier, animals accepted donor-specific ACI skin and primarily vascularized cardiac allografts permanently, while rejecting third party Brown Norway (BN) grafts. The tolerance induced was also donor-specific in vitro as evidenced by specific hyporeactivity to the allogeneic donor lymphoid elements, yet normal reactivity to MHC-disparate third party rat lymphoid cells. This model for mixed chimerism in the rat will be advantageous to investigate specific transplantation tolerance to primarily vascularized solid organ grafts that can be performed with relative ease in the rat, but not in the mouse, and may provide a method to study the potential existence of organ- or tissue-specific alloantigens in primarily vascularized solid organ allografts.

Analysis of early reconstitution events in the SCID mouse thymus following rat bone marrow cell transplantation.

In this work, we provide comprehensive evidence that sublethally irradiated Thy-1.2 SCID mice can be used as a model system for thymus homing and reconstitution after intravenous transfer of rat bone marrow cells. Full short-term SCID mouse thymus reconstitutions were obtained using a plastic nonadherent low-density rat bone marrow cell subset. Cell counts and flow cytometric analysis showed that at 3 weeks post-transfer the SCID mouse thymus contained up to 41 x 10(6) Thy-1.1high rat lymphoid cells comprising the expected percentages and distribution of CD2+, CD5+, CD3+, alpha beta TCR+ and CD4+ CD8+ cells. As seen on cryostat sections, bone marrow-derived MHC class II+ accessory cells had already developed by 2 weeks post-transfer, preceding the thymocyte expansion apparent at 3 weeks. Thus, analysis of the early events of SCID thymus reconstitution by rat bone marrow cells shows that they closely resemble those described in congenic animals and points out the temporally distinct development of dendritic cells and thymocytes. The SCID mouse-rat chimera model system represents a new in vivo tool for manipulating rat T-cell differentiation from bone marrow-resident precursor cells and in addition supports our previous xenogeneic reconstitution studies performed in organ culture.

Histamine releasing activity and its inhibition by supernatants from cultured lymphoid cells of Nippostrongylus brasiliensis-infected rats.

The influence of Nippostrongylus brasiliensis infection on the capability of HRF (Histamine Releasing Factor) generation by rat lymphoid cells in vitro has been studied. Spleen cells and thymocytes of normal and Nippostrongylus brasiliensis-infected rats were cultured in the presence of nonspecific mitogen (PHA) or specific N. brasiliensis antigen (NbAg), and cell-free supernatants fractionated by Sephadex G-75 chromatography were tested on homologous mast cells for histamine releasing activity. The results show that PHA-stimulated lymphoid cells from both normal and infected rats produced a factor releasing histamine from mast cells. Histamine releasing activity was not detected when lymphoid cells of N. brasiliensis-infected rats were cultured in the presence of NbAg. Moreover, supernatants of these cultures diminished HRF-induced histamine release from mast cells, suggesting the production of factor(s) inhibiting this release. This histamine release inhibiting activity was detected in fractions in Sephadex G-75 chromatography of supernatants from the cultures of NbAg-stimulated thymocytes of infected rats.

A selective culture system for generating terminal deoxynucleotidyl transferase-positive lymphoid cells in vitro. V. Detection of stage-specific pro-B-cell stimulating activity in medium conditioned by mouse bone marrow stromal cells.

The selective in vitro generation of rat, mouse, and human terminal deoxynucleotidyl transferase-positive (TdT+ lymphoid cells in our long-term xenogeneic bone marrow (BM) culture system is characterized by physical interaction between the developing lymphocytes and mouse BM-adherent stromal cells and macrophages. In the present study, experiments in which micropor)us membrane culture inserts were inoculated with rat BM cells demonstrated that although the generation of primitive B-lineage lymphoid cells requires the presence of a mouse BM feeder layer, cognitive recognition events are not necessary. Similarly, cell-free (and serum-free) medium conditioned with mouse BM (but not thymus or spleen) adherent cells and stromal-cell lines therefrom supported the proliferation of early rat lymphoid cells in a dose-dependent manner. Double immunofluorescence for incorporated bromo-deoxyuridine (BrdU) and early B-lineage markers of rat BM lymphoid cells maintained in culture inserts or conditioned medium (CM), and studies of their in vitro and in vivo developmental potentials, indicated that the lymphoproliferative response resulted from the selective stimulation of lymphoid stem and/or progenitor cells. The most primitive of these target cells had a HIS24+ HIS50- TdT- cμ- sIg-, pre-pro-B-cell phenotype. Whereas this subset normally constitutes less than 2% of B-lineage BM cells in vivo, it comprises more than 25% of total lymphoid cells in vitro. In addition, the number of TdT+ cells, predominantly of the early pro-B-cell phenotype (HIS24+ HIS50- TdT- cμ- sIg-), was increased approximately tenfold above input levels. Based on these and previous findings, a schematic model is proposed for the developmental pathway of early B-lineage cells in rat BM from the level of the committed (possibly common) lymphoid stem cell to that of the pre-B-cell.

Phenotypic progression of a rat lymphoid cell line immortalized by human T-lymphotropic virus type I to induce lymphoma/leukemia-like disease in rats.

Rat lymphoid cells, TARS-1, immortalized by coculture with adult T-cell leukemia cells, were intraperitoneally injected into 65 newborn, inbred WKAH/Hkm rats. In most of the rats, tumor nodules were discernible 7 to 15 days after transplantation but were completely rejected within 5 to 6 weeks. Two rats with no tumor nodules exhibited gait disturbances and paralysis of the hind legs 3 to 4 weeks after transplantation. Histological and hematological examinations revealed that a lymphoma/leukemia-like disease had developed in one of the two rats, and the T-lymphoid cell line WLeuk-1 was established from peripheral blood mononuclear cells from this rat. When the WLeuk-1 cells were transplanted into newborn WKAH/Hkm rats, the animals died of a lymphoma/leukemia-like disease within several weeks after transplantation, in contrast to their rejection of the TARS-1 cells. Southern blot and karyotype analyses revealed that WLeuk-1 cells had retained the marker chromosomes and human T-lymphotropic virus type I (HTLV-I) integration patterns of the parent cell line, TARS-1. The additional specific chromosome abnormalities 3p+,t (12;13), and Xq+ were found in the WLeuk-1 cells. Moreover, the expression of HTLV-I structural proteins was slightly depressed in WLeuk-1 cells, while that of the transacting factors p40tax and p21x, but not that of p27rex, was enhanced about fivefold compared with that in TARS-1. The transactivating function of p40tax was intact in WLeuk-1, as evidenced by enhanced interleukin-2 receptor alpha chain expression. These results suggest that aberrant expression of HTLV-I regulatory genes and alteration of cellular genes were associated with the phenotypic progression of the WLeuk-1 cell line.

Involvement of dendritic cells in early insulitis of BB rats

In this study, subsets of mononuclear infiltrates in pancreatic islets of BB rats at different stages of insulitis were determined by various monoclonal antibodies against rat lymphoid cells, including a mouse monoclonal IgM antibody that distinguishes between macrophages and dendritic cells (mAb 1F119). 1F119 + dendritic cells were absent in and around islets of Wistar control rats. In BB rats, the first alteration of islets detectable by immunohistochemistry when compared with normal islets was the enhanced expression of 1F119 antigen around and in the islets (17% 1F119 + islets). At disease stage 1 (i.e. no leukocyte infiltration after HE staining), lymphocytes and macrophages were almost absent. At disease stage 2 (leukocyte infiltration <20 cells), a more intense form of dendritic cell infiltration was seen (stage 1 versus stage 2, P <0.0001). In addition, ED2 + and ED3 + cells were present around the islets (50% of islets were infiltrated with 1F119 + cells versus 16% with ED2 +, 19% with ED3 +, 11% with W3 25 + cells, and 11% with OX8 + cells, P <0.0001). At disease stage 3 (>20 cells), a clear increase of ED2 + and ED3 + macrophages and of W3 25 + and OX8 + T-lymphocytes in the infiltrates was observed. These observations suggest a role for antigen presenting dendritic cells in the initiation of immune reaction in type 1 diabetes of BB rats.

MIXED XENOGENEIC CHIMERAS (RAT + MOUSE TO MOUSE)

We report the induction of stable and reliably detectable mixed xenogeneic chimerism through the coadministration of a mixture of untreated rat bone marrow plus T cell-depleted mouse bone marrow into B10 recipients conditioned with total body irradiation (TCD B10 mouse + untreated F344 rat----B10 mouse). Recipients repopulated as true mixed lymphopoietic chimeras, with from 1-21.6% rat-derived lymphoid cells in peripheral blood and splenic lymphoid tissue. Production of rat platelets was also demonstrated. Rat platelet and lymphoid chimerism was reliably detectable in chimeras from 1 to 7 months following reconstitution, suggesting engraftment of the rat bone marrow stem cell. Production of each stem cell-derived lineage appeared to be under independent regulation since a significantly greater proportion of platelets were rat-derived (24-81%) than were lymphocytes (1-21.6% rat), while erythrocytes were preferentially syngeneic (less than 2% rat). The tolerance induced by this model was highly donor strain-specific: donor-specific rat and mouse skin grafts were accepted while MHC-disparate third-party mouse (C3H; H-2k) and rat (Wistar Furth; Rt1Au) skin grafts were promptly rejected. Although specifically prolonged xenogeneic donor rat skin grafts underwent a slow chronic rejection, and some totally disappeared. In spite of this, chimeras retained their lymphoid chimerism, suggesting the presence of skin-specific antigens. This model for mixed xenogeneic chimerism with reliably detectable rat lymphoid cells may provide a model to study the existence of tissue-specific antigens across a species barrier, as well as mechanisms responsible for the induction and maintenance of this strain-specific transplantation tolerance.

Study on the lipid composition of aging fischer‐344 rat lymphoid cells: Effect of long‐term calorie restriction

Long-term calorie restriction (LCR) is widely known to increase the survival rate of laboratory rodents and appears to retard the aging and senescence process. The present study was undertaken in Fischer-344 male rats maintained on ad libitum (AL) or LCR (40% less food intake than AL starting at 6 weeks of age). Age-associated changes in the proliferative response of lymphoid cells to mitogenic stimuli were studied in relation to alterations in the fatty acid composition of adherent and non-adherent-enriched subpopulations of spleen cells. Increases in spleen cell long-chain highly unsaturated fatty acids (20:4, 22:4 and 22:5) were accompanied by decreases in linoleic acid (18:2) in aging AL-fed rats. However, LCR stabilized levels of 18:2 and prevented the rise in highly unsaturated fatty acids. In addition, LCR markedly modulated the fatty acid profiles of thymocytes and bone marrow cells. A 70% decline in concanavalin A (Con A) stimulated [3H]thymidine uptake of spleen cells from AL animals was normalized by LCR. Splenic reduced glutathione (GSH), a potential modulator of the mitogenic response, was unaffected by age and nutritional regimen. Thus, normalization of lymphoid cell fatty acid composition by LCR parallels the preservation of mitogenic responsiveness to Con A.