META-controlled env-initiated transcripts encoding superantigens of murine Mtv29 and Mtv7 and their possible role in B cell lymphomagenesis.

Spontaneous germinal center (GC)-derived B cell lymphomas of SJL mice (RCS) transcribe a 1.8-kb Mtv-29 mRNA under control of the META-env promoter. The encoded vSAg29 stimulates syngeneic Vbeta16(+) CD4(+) T cells, thereby acquiring T cell help necessary for RCS growth. Other strains of B cell lymphoma-prone mice include Mtv29(+) C57L and MA/MyJ, and the Mtv29(-) Mtv7(+)-recombinant inbred strain, SW x J-1. The lymphomas of these mice produce similar mouse mtv-vSAg-encoding mRNA, as characterized by Northern blotting, PCR, and RNase protection. A 1.8-kb mRNA in C57L/J and MA/MyJ lymphomas hybridized with an Mtv29-specific oligonucleotide, whereas SW x J-1 lymphomas produced 1.8-kb transcripts hybridizing with an Mtv7-specific oligonucleotide. Similar META-env-initiated transcripts were absent from LPS-activated B cells from any strain examined but were detected in Peyer's patch RNA from SJL mice. Like typical SJL-derived RCS, all these lymphomas stimulated syngeneic CD4(+) T cells and Vbeta16(+) T hybridoma cells. Immunohistochemical staining of primary tumors showed the presence of peanut agglutinin binding (PNA(+)) highly mitotic lymphoblasts, suggesting their GC derivation. The findings indicate that this novel mRNA for Mtv29 is present in B cell lymphomas from several Mtv29(+) mouse strains. Additionally, this is the first description of the ability of Mtv7 to produce transcripts that are controlled and spliced identically to those of Mtv29 and that are expressed in SW x J-1, I-A(s+), lymphomas that also stimulate Vbeta16(+) T cells. Our results suggest an important role for mouse mtv-vSAgs and Vbeta16 T cell stimulation in the development of GC-derived murine B cell lymphomas.

Requirement for reverse immune surveillance for the growth of germinal center-derived murine lymphomas.

The concept of reverse immune surveillance, first conceived over 12 years ago, described the relationship that existed between germinal center-derived B cell lymphoma cells and the host immune system in SjL/J mice. According to reverse immune surveillance, recognition of tumor cell antigens and a response by the host immune system is required for tumor growth. The phenomenon of reverse immune surveillance related to B cell lymphomas has recently also been characterized in another inbred mouse strain, C57L/J. Moreover, elements of reverse immune surveillance have been observed in several other mouse strains that develop B cell lymphomas, suggesting that this lymphomagenic mechanism may be more common than first envisioned. In SJL and C57L mice, the B lymphoma cells express an MMTV-encoded superantigen (vSAg29) that stimulates syngeneic CD4+ T cells bearing Vbeta16 in their TCR. In contrast to the mRNAs for other MMTVs in normal mouse B cells, vSAg29 mRNA initiates in the env (META) region, undergoes splicing in the 3' env region, and continues through the 3' LTR. Copious cytokine production, including IFN-gamma, IL-4 and IL-5 accompanies the response of the T cells to this vSAg. In addition to cytokines produced by vSAg-responsive T cells, more recent evidence indicates that another cytokine, LTalphabeta2, which is expressed on the lymphoma cell surface, also plays a role in the promotion of the B cell lymphoma growth. It is possible that interaction with LTbeta-R on follicular dendritic cells or other stromal elements facilitates tumor growth by preventing apoptosis of the malignant B cells. To what degree these findings in the mouse are relevant to the development and/or growth of human B lymphoma cells remains to be determined. However, endogenous retroviral sequences do exist in the human genome. Interestingly, some of these sequences are homologous to MMTV, and are transcribed in B lymphoblastoid cells. Moreover microorganisms that are infectious for human B cells, such as EBV and Herpes Virus 8, may also produce superantigens.

B cell lymphomas of C57L/J mice; the role of natural killer cells and T helper cells in lymphoma development and growth.

The Hodgkin's-like Type B neoplasms which arise spontaneously in aging C57L mice (25% incidence at 21 months of age) were first reported over 40 years ago, but since then relatively little has been published about these lymphomas. Based on previous studies in SJL mice, we investigated the phenotypic and functional properties of C57L-derived lymphomas in relation to Mtv29-encoded vSAg expression by the tumor cells, and their ability to stimulate TCR Vbeta-restricted T cells. The cell surface phenotype of the C57L lymphomas indicates a B cell origin (sIg(+), MHC II(+)). These B lymphoma cells also express co-stimulatory molecules [B7-1 (CD80) and HSA (CD24)], and stimulate marked proliferation of syngeneic CD4(+) T cells. C57L B lymphoma cells exhibit Mtv-encoded mRNA by northern analysis, and also stimulate IL-2 production from Vbeta16(+) T cell hybrids, suggesting a role for Mtv 29 in this syngeneic T cell response. After transfer to syngeneic recipients, primary C57L lymphomas grow slowly, if at all. However, tumor growth is greatly accelerated by pretreatment of C57L recipients with anti-asialo GM1 antibody (but not anti-CD8 mAb), suggesting that NK cells play a major role in inhibiting lymphoma growth. If, in addition to anti-asialo GM1, the mice are also pretreated with anti-CD4 mAb, tumor growth is markedly inhibited, indicating that the lymphoma-responsive syngeneic CD4(+) T cells promote tumor growth. Therefore, although the vSAg-induced response stimulated by vSAg29 expressing lymphoma cells in syngeneic TCR Vbeta-restricted CD4(+) T cells is an important etiologic factor in this type of B cell neoplasm both in C57L and in SJL mice, the final outcome of the spontaneous neoplastic process appears strongly influenced by endogenous NK activity in aging mice.

Diverse fine specificity and receptor repertoire of T cells reactive to the major VP1 epitope (VP1230-250) of Theiler's virus: V beta restriction correlates with T cell recognition of the c-terminal residue.

Theiler's murine encephalomyelitis virus induces chronic demyelinating disease in genetically susceptible mice. The histopathological and immunological manifestation of the disease closely resembles human multiple sclerosis, and, thus, this system serves as a relevant infectious model for multiple sclerosis. The pathogenesis of demyelination appears to be mediated by the inflammatory Th1 response to viral epitopes. In this study, T cell repertoire reactive to the major pathogenic VP1 epitope region (VP1233-250) was analyzed. Diverse minimal T cell epitopes were found within this region, and yet close to 50% of the VP1-reactive T cell hybridomas used V beta 16. The majority (8/11) of the V beta 16+ T cells required the C-terminal amino acid residue on the epitope, valine at position 245, and every T cell hybridoma recognizing this C-terminal residue expressed V beta 16. However, the complementarity-determining region 3 sequences of the V beta 16+ T cell hybridomas were markedly heterogeneous. In contrast, such a restriction was not found in the V alpha usage. Only restricted residues at this C-terminal position allowed for T cell activation, suggesting that V beta 16 may recognize this terminal residue. Further functional competition analysis for TCR and MHC class II-contacting residues indicate that many different residues can be involved in the class II and/or TCR binding depending on the T cell population, even if they recognize the identical minimal epitope region. Thus, recognition of the C-terminal residue of a minimal T cell epitope may associate with a particular V beta (but not V alpha) subfamily-specific sequence, resulting in a highly restricted V beta repertoire of the epitope-specific T cells.

The feasibility of using blood bank-stored (4 degrees C) cord blood, unmatched for HLA for marrow transplantation.

The main purpose for storing large numbers of umbilical cord blood (CB) units by cryopreservation is to obtain a close HLA match for use in bone marrow transplantation. The use of partially matched or unmatched CB has been suggested, and publications about the success of 3 antigen mismatches have given some credence to this suggestion. Graft vs host disease still is considered a major barrier for successful CB transplantation. The cost per frozen CB unit of approximately $15,000 considerably limits its availability in developing countries. Eleven human umbilical cord specimens were stored in gas-permeable bags at 4 degrees C for up to 3 weeks. Clonal growth, replating efficiency in methylcellulose cultures, differential count, and flow cytometric immunophenotyping results were examined at intervals up to 21 days. Mixed lymphocyte cultures were evaluated on 13 similarly stored specimens at intervals up to 14 days. When plated at 1, 10, and 21 days, the combined percentage of the more primitive colonies increased on days 10 and 21. Replating efficiency of blast cell colonies when stem cell factor was added was 81.2% and 67.8% on days 10 and 21, respectively. When mononuclear cells were immunophenotyped, the mean percentage of CD34+ and CD117+ cells, considered primitive stem cell markers, increased significantly from day 1 to day 21. The ability of stored CB cells to respond to phytohemagglutinin or alloantigens decreased progressively from day 1 to day 14. By day 14, the reactivity of CB responder cells, in mixed lymphocyte cultures, to fresh allogeneic CB stimulator cells declined significantly. These findings suggest that CB can be stored in existing blood bank facilities and retain its hematopoietic potential for transplantation. Furthermore, it may be feasible to combine individual CB samples to provide a sufficient number of viable stem cells for transplantation, substantially expanding the number of potential recipients.

Effects of cord blood transfer on the hematopoietic recovery following sublethal irradiation in MRL lpr/lpr mice.

The potential of cord blood (CB) to serve as a rich source of stem cells and stem cell factors is receiving increasing attention. In addition, perhaps because of the early ontogeny of these cells or the lack of surface antigens, cord blood stem cells do not appear to require close identity with the recipient. In the present pilot study, we investigated the presence of a hematopoiesis enhancing effect (HEE) by assaying the ability of human cord blood cells to augment hematopoiesis across a species barrier. For these experiments, autoimmune-prone MRL-Ipr/Ipr mice were exposed to sublethal levels of irradiation and cord blood administration to study the role of factors present in human cord blood in augmenting the rate of lymphopoiesis. This strain was chosen because of the increased presence of peripheral T and B subpopulations, namely the B-1 and CD4/CD8 double negative T-cell subpopulations, which do not arise directly from bone marrow precursors, but rather accumulate with age. MRL-Ipr/Ipr mice were sublethally irradiated and reconstituted with syngeneic bone marrow (BM) cells or with human cord blood cells or peripheral blood mononuclear cells (PBMC), or were left unreconstituted. At 2 weeks post-treatment, lymphoid populations in the spleen and lymph nodes were studied as a measure of hematopoiesis. Factors present in cord blood were able to augment hematopoiesis over that which occurred endogenously. At 2 weeks postirradiation, recipients of BM cells displayed the fastest rate of peripheral lymphoid recovery, nonreconstituted mice showed the slowest lymphoid recovery, and recipients of cord blood recovered their lymphoid populations at an intermediate rate. Similarly, myelopoiesis was increased in irradiated SJL/J recipients of human cord blood. Thus, human cord blood cells appear to produce/induce factors that may act as an adjunct to increase stem-cell activity.

Endogenous hematopoietic reconstitution induced by human umbilical cord blood cells in immunocompromised mice: implications for adoptive therapy.

Human umbilical cord blood (HUCB) cells show promising advantages over bone marrow (BM) cells for a variety of diseases that require transplantation. We observed that lethally irradiated SJL/J mice given a single injection of HUCB cells survive, whereas vehicle-injected mice do not. Because survival is not due to long-term engraftment of HUCB cells, we used this HUCB/mouse model to investigate additional therapeutic benefits of HUCB cells. We investigated the mechanism by which HUCB cells accelerated endogenous hematopoiesis in mice that received either lethal (9.5 Gy) or lower-dose (8.0 Gy) radiation and then were given a single injection of HUCB mononuclear cells. Compared to irradiated control mice, the lethally irradiated, HUCB-injected group showed significant increases in peripheral white blood cell counts, red blood cell indices, and granulocyte-macrophage colony-forming units (CFU-GM) by 3 weeks. In contrast, no significant differences in these parameters were observed between control and HUCB-injected mice that received the lower dose of irradiation. Moreover, regardless of the radiation dose, only HUCB-injected mice exhibited immune responses comparable to those of age-matched normal mice. The clinical relevance of these observations was determined in long-term, culture-initiating cell assays with human BM stem cells and irradiated (gamma-) HUCB cells. CFU-GM colonies were detectable in cultures containing gamma-HUCB cells by day 15, but were undetectable in cultures without gamma-HUCB cells until day 40, suggesting a hematopoietic stimulatory role for HUCB cells. Overall, the results indicate that in addition to their use for transplantation, HUCB cells also may be used as an adjuvant therapy to enhance hematopoietic reconstitution and immunocompetence of the host. This hematopoiesis-enhancing effect represents a heretofore unrecognized function of HUCB cells.

Long-term exposure of HL60 cells to 1,25-dihydroxyvitamin D3 reduces their tumorigenicity: a model for cancer chemoprevention.

Several lines of evidence suggest that 1,25-dihydroxyvitamin D3 (1,25D3) may be important in chemoprevention of human cancer. Here, we show that human promyelocytic leukemia cells HL60 cultured in the presence of 30 nM 1,25D3 (30A cells) for 3 years exhibited a reduced rate of tumor growth when injected into nu/nu mice, while cells grown in 40 nM 1,25D3 (40AF cells) failed to form detectable tumors in 11 out of the 12 inoculated mice, interestingly, both 30A and 40AF cells grew approximately twice as fast as the parental HL60-G cells under tissue culture conditions, even in the presence of 1,25D3, to which they developed resistance. Tests of the susceptibility of these cells to natural killer (NK) cell cytotoxicity showed that 40AF, but not HL60-G or 30A cells, were targets for the murine spleen NK cells. However, lysis of 30A cells was also detected when human NK cells were used in this assay, though the effector-to-target cell ratio necessary to obtain significant lysis above background levels was higher for 30A (80:1) than for 40AF (10:1) cells. These results suggest a mechanism for the reported chemopreventive effects of sunlight-generated 1,25D3 or dietary vitamin D3.

The effect of human cord blood on SJL/J mice after chemoablation and irradiation and its possible clinical significance.

There is evidence from the existing published literature that human umbilical cord blood, when used for purposes of bone marrow transplantation, does not necessarily have to be HLA matched in order to be efficacious. These reports include experimental observations on the ability of human umbilical cord blood to rescue lethally irradiated mice and clinical observations from China wherein HLA mismatched umbilical cord blood has been engrafted successfully in children with malignant disease. The study reported herein describes an experimental immunocompetent murine model to determine if human umbilical cord blood can be used to improve survival after chemoablation and irradiation. The animals received chemoablation followed by irradiation, and irradiation alone. The presence of human DNA in these mice following injection of human umbilical cord blood cells was determined, and the immunological status of the animals was evaluated. Animals receiving human umbilical cord blood cells after chemoablation and irradiation had a better mean survival at day 50 than animals receiving syngeneic marrow. Human DNA could be found in various organs, particularly the lung, spleen and liver of the mice for the first 30 days. Thereafter, human DNA became more difficult to detect but trace amounts of human DNA could be found up to one year later. The results of mixed lymphocyte reactions and phenotype analyses for murine T cell markers performed after injection of HUCB cells both indicated endogenous repopulation, and relatively intact immune systems in these mice. Since human umbilical cord blood allowed mice to survive the lethal effects of chemoablation plus irradiation, or irradiation alone, with reconstitution of the animals' own, relatively intact, immune systems, it would appear that HLA mismatched human umbilical cord blood could potentially be used as an adjuvant treatment for patients with advanced malignancies or other diseases for which hematopoietic reconstitution is indicated.

IL-10 production in a CD5+ B cell lymphoma arising in a CD4 monoclonal antibody-treated SJL mouse.

A majority of SJL mice develop spontaneous reticulum cell sarcomas (RCS) at about 1 year of age which can be transplanted into young SJL recipients. Previous studies have shown that RCS tumors are of B cell lineage, and that the development of these lymphomas and their subsequent growth depends upon host-derived T helper cell factors. In vivo treatment of SJL mice with anti-CD4 monoclonal antibody (mAb) prevents the development of the characteristic B lymphomas. Most of the mAb-treated animals were tumor free and had a significantly prolonged life span. However, one such CD4 mAb-treated mouse developed a transplantable IgM+ CD5+ B cell lymphoma (designated NJ101), which has not previously been described in SJL/J mice. NJ101 is clonal on the basis of a discrete non-germ line Ig heavy chain gene rearrangement by Southern blot analysis. Unlike the sIg- CD5- transplantable RCS-X cell line, the IgM+ CD5+ NJ101 lymphoma cells will grow in immuno-compromised hosts, such as irradiated recipients or in recipients treated with CD4 mAb in vivo. The RCS (B cell) lymphoma requires CD4+ T cells for progressive growth, whereas the growth of the CD5+ B lymphoma cells is enhanced by the removal of such cells. Thus, CD5+ B cell clonal development may be aided by the removal of regulatory T cells and/or the malignant CD5+ B cells may produce their own growth factors in an autocrine manner. Examination of IL-10 message by quantitative polymerase chain reaction techniques indicate that the CD5+ B lymphoma cells produce increased levels of IL-10 relative to normal LN cells or purified RCS lymphoma cells. These results suggest that two different types of B cell tumors, both of which can develop in SJL mice, have different growth requirements. Furthermore, treatment to prevent the occurrence of the characteristic RCS malignancy of SJL mice may lead to the development of another form of B cell neoplasia.

Murine survival of lethal irradiation with the use of human umbilical cord blood.

We have found that human umbilical cord blood (HUCB) will routinely protect mice exposed to lethal levels of irradiation. At the end of 50 days, over seventy percent (70%) of mice injected with HUCB survived 900 cGy or irradiation, which produced 100% deaths in the uninjected control animals. Moreover, there was some evidence that human colony stimulating factors further improved survival. Anti-Natural Killer cell (NK) antibody was utilized along with HUCB in these studies, however, Anti-NK cell serum alone had no radioprotective effect in mice. The studies reported here suggest the possibility of utilizing HUCB for immediate protection of humans from lethal irradiation.

Syngeneic B lymphoma cells provide a unique stimulus to natural killer (NK) cells in genetically low-NK SJL/J mice.

SJL/J mice are a genetically low-NK strain, and their cytotoxic activity cannot be augmented with conventional NK inducers. In contrast, effector cells taken from the lymphoid tissues of SJL mice bearing a syngeneic B cell lymphoma (RCS) show variable, but significant levels of cytotoxic activity against NK-susceptible targets, such as YAC-1. Previous results suggested that the RCS cells themselves contributed to this cytotoxicity. However, results presented here indicate the most, if not all of the activity present within the lymphoid tissues of RCS-bearing mice is mediated by RCS-activated, host NK cells. These results were confirmed by in vitro studies, which demonstrate that both gamma irradiated (gamma-) RCS cells and gamma-allogeneic spleen cells induce cytotoxic activity in SJL spleen cells against YAC targets. However, the cytotoxicity induced by gamma-allogeneic cells is mediated largely by lymphokine-activated killer (LAK) cells, since these effectors also lyse NK-resistant target cells, such as L1210. In contrast, the cytotoxic effector cells that are induced by syngeneic gamma-RCS cells cause lysis of YAC targets, but not L1210 target cells. These data indicate that the syngeneic B cell lymphomas of SJL mice are a unique stimulus for host NK cells in this strain. Since activated NK cells produce a variety of lymphokines, RCS stimulation of host NK cells in SJL mice may provide some of the growth-promoting lymphokines that are known to be necessary for progressive growth of these lymphoma cells.

Paraproteins and primary lymphoma in SJL mice. II. Primary lymphomas do not produce paraprotein.

The incidences of Ig paraprotein (PP) and reticulum cell sarcomas (lymphomas) were studied in a group of 75 SJL mice, 9-11 months of age. PP and lymphoma were observed in 52% of the mice. PP alone was observed in an additional 27% and lymphoma alone in an additional 11% of mice. In attempts to establish a causal relationship between lymphomas and PP, three approaches were used: (a) Serum PP levels were followed during lymphoma growth in primary lymphoma bearing mice and found to decrease rather than to increase. (b) Recipients of primary transplants were examined for appearance of PP-related idiotypes (Id) in their sera and for lymphoma growth. The Id appeared early (Day 10-11) and then disappeared, while lymphoma growth usually was detectable by greater than or equal to 1 month. (c) One of the primary lymphomas was propagated as a tissue culture line and found not to produce any PP or intact Ig. It is concluded that the PP of SJL mice are not produced by their lymphomas. Other possible relationships are discussed, including the role of the PP as a symptom of a prelymphomatous stage that develops in a very high percentage of SJL mice.

Influence of T-helper cell specific monoclonal antibody on progressive growth of B-cell lymphomas in SJL/J mice: correlation of acute treatment dosage with tumor dormancy or complete remission in long-term survivors.

Transplantable follicular center cell lymphomas of SJL/J mice (SJL/FCC) are B-cell-derived tumors that stimulate host T-helper (TH) cells and grow progressively in the lymphoid tissues of immunocompetent, syngeneic recipients. The host TH cells that respond to the I-As determinants on SJL/FCC lymphoma cells produce a variety of lymphokines, some of which (e.g., IL-5) promote in vitro tumor growth. The results presented here demonstrate that removal of the cellular source of tumor growth-promoting lymphokines by treatment of lymphoma-injected mice with TH cell specific monoclonal antibodies (anti-L3T4a) inhibits progressive tumor growth and prolongs survival significantly. However, long-term survival is mediated by different mechanisms, depending on the dosage of L3T4a mAb used. Tumor cells are present, but dormant, in mice that receive low-dose (less than 200 micrograms/injection) treatment. In contrast, tumor cells are undetectable in mice that receive high dose (1200 micrograms/injection) treatment.

Production of human to mouse xenografts by umbilical cord blood.

Utilizing human umbilical cord blood, it has been possible to create in irradiated animals a human to mouse xenograft. To facilitate hematopoietic reconstitution, SJL/J mice, which are functionally low in natural killer (NK) cells, were treated with anti-Asialo GM1 antibodies (anti-NK) and irradiation prior to injection of cord blood mononuclear cells. In contrast, SJL/J mice with the "beige" (bg/bg) mutation, which confers a functional NK cell deficiency, required only irradiation for successful transplantation. Human cells, detected by means of DNA probes, were demonstrated in the lungs and lymph nodes of irradiated animals up to 6 months after injection of the human cord blood cells.

Reticulum cell sarcomas of SJL mice have rearranged immunoglobulin heavy and light chain genes.

The immunoglobulin (Ig) heavy (H) and light (L) chain gene rearrangements of the high incidence SJL lymphomas (reticulum cell sarcoma, RCS) have been analyzed. Both primary and transplanted RCS show rearrangements of H and kappa L chains, demonstrating that these tumors are of B cell origin. These data are consistent with previous results indicating that these tumors are a mouse model for follicular lymphoma. A long-term transplanted line and the in vitro line derived from it, cRCS-X, have a single rearranged JH-C gamma 2a fragment and one rearranged C alpha gene fragment which does not hybridize with a probe for the JH gene segments. These cell lines also have two rearranged J kappa-C kappa fragments. Primary tumors and early passages are more heterogeneous with respect to Ig gene rearrangements, possibly because more than one B cell clone is present. Although no synthesis of IgG2a, or of any Ig, could be detected by the in vitro cRCS-X cells, these cells contain abundant poly(A)+ RNA that hybridize with gamma 2a and kappa probes as well as lesser amounts of alpha and epsilon RNA. None of these H chain RNA hybridized with probes for the JH gene segments. The epsilon and alpha RNA are the same size as transcripts of germ-line CH genes which have been identified in other systems. However, the gamma 2a RNA are smaller than previously described germ-line C gamma 2a RNA and appear to be transcribed from aberrantly rearranged JH-C gamma 2a genes.

T-helper-cell-specific monoclonal antibody inhibits growth of B-cell lymphomas in syngeneic SJL/J mice.

Transplantable follicular center cell lymphomas of SJL/J mice are B-cell tumors that stimulate proliferation of host T-helper (TH) cells and which grow progressively in the peripheral lymphoid tissues of immunocompetent recipients. However, tumor growth is compromised in immunosuppressed syngeneic recipients, suggesting that the host response to SJL follicular center cell (SJL/FCC) lymphoma cells is required for optimal tumor growth. In vitro studies indicate that the host TH cells (Lyt-1+, 2-, L3T4a+) which respond to the major histocompatibility complex (MHC) class II (I-As) surface determinants on the SJL/FCC lymphoma cells produce a variety of lymphokines, some of which may promote tumor growth in vivo. The results of this study demonstrate that treatment of lymphoma-injected mice with L3T4a-specific mAb inhibits the growth of the SJL/FCC lymphoma cells, despite the fact that these tumor cells do not express L3T4a determinants. Thus, in this model, mAb therapy targeting host immune cells rather than the tumor cells is an effective means to control tumor growth. Long-term observation of SJL/FCC lymphoma-injected, anti-L3T4a mAb-treated mice reveals prolonged survival of the majority of these animals with periodic recurrence of tumor growth. During periods of remission, LN cells from these long-term surviving animals were unable to mount the characteristic in vitro host response to irradiated SJL/FCC lymphoma cells. These results provide direct evidence that SJL/FCC lymphoma cells fail to retain their characteristic neoplastic properties in a microenvironment that is initially devoid of tumor-responsive TH cells.

Restriction of tumor growth in mice by sodium-deficient diet.

Generalized malnutrition results in inhibition of tumorigenesis and tumor growth in experimental animal models. Neither the specific nutrient deficiency nor the mechanism has been definitely elucidated. We have shown previously that dietary sodium deprivation in rapidly growing rats retards protoplasmic growth. This effect was correlated to the extracellular fluid (ECF) volume expansion which is dependent on sodium accumulation. Since solid tumors are composed of a large quantity of ECF (which includes plasma volume) it was postulated that preventing the accumulation of new ECF by means of sodium restriction would influence tumor growth. The present study was designed to determine the effects of salt restriction on tumor growth and to relate these effects to ECF volume. Approximately 10(6) viable B16 melanoma cells were injected into C57BL/6 x DBA/2 F1 and C57 mice. A salt restricted diet (sodium less than 3 microeq/g) was provided ad libitum. The drinking solution was distilled water for the experimental group and 0.45% saline solution for the controls. There was a significant decrease in tumor growth rates during sodium restriction. The total body ECF volume increased when dietary sodium was supplied but did not change during salt restriction. Therefore, the only source for the ECF in the tumor mass was from nontumorous tissue. We conclude that during dietary sodium restriction solid tumor growth is retarded and can proceed only to the extent that ECF is released from cachectic body tissues.