Neutrophils and monocytes transport tumor cell antigens from the peritoneal cavity to secondary lymphoid tissues

Antigen-transporting cells take up pathogens, and then migrate from sites of inflammation to secondary lymphoid tissues to induce an immune response. Among antigen-transporting cells, dendritic cells (DCs) are believed to be the most potent and professional antigen-presenting cells that can stimulate naïve T cells. However, the cells that transport antigens, tumor cell antigens in particular, have not been clearly identified. In this study we have analyzed what types of cells transport tumor cell antigens to secondary lymphoid tissues. We show that neutrophils, monocytes and macrophages but not DCs engulf X-irradiated P388 leukemic cells after their injection into the peritoneal cavity, and that neutrophils and monocytes but not macrophages migrate to the parathymic lymph nodes (pLN), the blood, and then the spleen. The monocytes in the pLN comprise Gr-1⁻ and Gr-1⁺ ones, and some of these cells express CD11c. Overall, this study demonstrates that neutrophils and monocytes transport tumor cell antigens from the peritoneal cavity to secondary lymphoid tissues.     

Specific triggering of macrophage accumulation at the site of secondary tumor challenge in mice with concomitant tumor immunity

Intraperitoneal injection of tumor cells triggers a greater influx of macrophages into the peritoneal cavity of mice with concomitant immunity than into control mice. The specificity of the reaction parallels the specificity of concomitant immunity. The newly arrived cells were classified as young monocyte-derived macrophages on the basis of morphology, position, and change in sedimentation velocity profiles, chemotaxis, and labeling in vivo after tritiated thymidine injection.     

DYNAMICS OF LEUKOCYTE MIGRATION INTO THE MOUSE ASCITES TUMOR

There is a marked increase in the number of peritoneal leukocytes (lymphocytes, monocytes and granulocytes) during the growth of Ehrlich ascites tumor in mice. No local proliferation (as indicated by a labeling at 1 hr following a single ³H-TdR injection) was observed in the normal peritoneal leukocytes or those in the ascites tumor, except for a very minor labeling of some tumor macrophages. Kinetics of peritoneal leukocytes was studied with a series of twelve injections of ³H-thymidine (20 μCi every 8 hr) in normal mice as well as mice injected with 10⁶ tumor cells i.p. 2 hr after the last ³H-TdR injection. Animals were sacrificed at intervals up to 6 days. Granulocyte labeling in the blood as well as peritoneal space was near 100% in both groups of animals at all the intervals. Temporal changes in the labeling of lymphocytes (from 10% at 0 day to 22% at day 6), and monocytes (from 20% at 0 day to 57% at day 6) were identical in the blood and peritoneal space of normal animals, indicating a free exchange of cells between these compartments. Higher labeling indices than those in the controls were attained in the blood of tumor-bearing hosts (viz 40% for lymphocytes and 80% for monocytes at 6 days) suggesting an increased turnover of these cells in the circulation. In addition, peritoneal mononuclear cells of tumor-bearing mice showed even a higher labeling than those in the blood (viz 65% for lymphocytes and 92% for monocytes at 6 days) indicating a selective migration and/or retention of newly formed cells within the tumor, in contrast to a random migration into the normal peritoneal cavity. Furthermore, an identical labeling of macrophages to that of monocytes within the tumor indicated a short monocyte-macrophage transition. The preferential accumulation of young mononuclear cells into the tumor may be of functional importance.     

Wound healing and angiogenic properties of supernatants from Lactobacillus cultures

Extracts or supernatants from cultures of Lactobacilli are used for their medicinal effects, including wound healing and immune system stimulating activity. We have studied the in vivo and in vitro effects of supernatants from bacterial cultures of two strains of Lactobacillus (LS) on tissue repair and angiogenesis. Subcutaneous injection of LS into rodent ears led to proliferation of blood vessels that also exhibited strong immunostaining for Flk-1 receptor. Some inflammatory cells were scattered among the blood vessels. The continuous influx of polymorphonuclear leukocytes (PMNs) and macrophages into transcutaneous wounds in mice treated with LS resulted in prolonged inflammatory phase of wound healing and delayed wound closure, including reepithelialization. Subcutaneous injection of Matrigel impregnated with LS into the abdominal wall led to rapid and transient influx of PMNs in the vicinity of the gel. LS stimulated the proliferation of murine macrophage J774.A1 cell line and porcine lymphocytes but not that of murine fibroblast AKR-2B cells. LS also induced production of TNF-alpha by J774.A1 cells and by porcine kidney epithelial LLC-PK1 cells. LS did not appear to have an effect on collagen production. In conclusion, our study demonstrates the potential of LS to function as a stimulator of the inflammatory stage of tissue repair, TNF-alpha production, and of angiogenesis.     

Immunologic approaches to the treatment of human cancer based on a guinea pig model

Summary Local immunotherapy is a form of cancer treatment where exogenous antigen is introduced into the area of the tumor. Under favorable circumstances, the perfused tumor regresses, systemic tumor-specific transplantation immunity is augmented, and distant microscopic metastases regress. Successful local immunotherapy requires an immunologically competent host, small tumor burden, and tumor located usually in the skin. A wide variety of biologic agents are capable of promoting local immunotherapy. BCG has been most widely studied. The antitumor activity of two different preparations of the Tice substrain of BCG were compared. No significant differences in antitumor activity were found. Alternative approaches to intralesional injection were sought. Intradermal injection of BCG adjacent to dermal tumors, prior to surgery, led to eradication of axillary metastases and to the development of tumor-specific transplantation immunity.Successful local BCG immunotherapy is a by-product of the host response to BCG infection. Involved are lymphocytes specifically sensitized to mycobacterial antigens, lymphocyte mediators, and macrophages which develop the capacity to kill tumor cells. Tumor-cell killing may be mediated by exocytosis of macrophage lysosomes into tumor cells. Complete and permanent tumor eradication probably requires the development of tumor-specific transplantation immunity mediated by sensitized lymphocytes. Local infection of the tumor may augment the development of this tumor-specific immunity.     

Characterization of guinea pig macrophages: I. Mobility and maturation of peritoneal cells following inflammatory stimuli

The wide spectrum of activities of macrophages may, in part, be a reflection of their different subpopulations. Sedimentation velocity was used in separate different guinea pig macrophage populations which were then compared in assays which measure some of the parameters of inflammatory responses. Normal resident, thioglycollate-induced exudates or cells elicited by ip injection of tuberculin into BCG-sensitized animals were studied. In thioglycollate exudates the small, possibly recently derived monocytes were most responsive to chemotactic agents whereas populations with high sedimentation velocity were more phagocytic, responsive to MIF and produced more LAF. By contrast, all macrophage populations or subpopulations from sensitized animals challenged with PPD were unresponsive to lymphocyte-derived chemotactic factor, endotoxin-activated serum, or to N-formyl-l-methionyl-l-phenylalanine in chemotaxis assays. In addition, macrophages from these animals did not migrate from capillary tubes in MIF assays. The lack of migration is presumably due to the action of MIF on these cells in vivo. Cells with low sedimentation velocity (predominantly lymphocytes), however, did migrate from capillary tubes. We conclude that small cells elicited by a nonspecific stimulus are probably attracted to the inflammatory site by chemotaxins and must subsequently enlarge or mature before they are able to respond to MIF, to be activated to produce LAF, or to exhibit extensive phagocytosis.     

Cellular mechanisms of tumor rejection in rats

The mechanisms of tumor rejection by cell-mediated immunity were reviewed in a rat autochthonous and syngeneic tumor-host system. The immune system could mediate a complete regression of autochthonous tumor if the tumor cells were immunogenic. Neutrophils and macrophages first appeared following transplantation of autochthonous tumor. Lymphocytes increased in the tumor tissue as the tumor began to show regression. Degenerated tumor tissue was infiltrated by macrophages and plasma cells. The identification of rat hematopoietic cells including various subsets of lymphocytes and inflammatory cells became possible owing to a variety of monoclonal antibodies reacting with these cells. Major populations of tumor-infiltrating lymphocytes (TIL) were found to be R1-3B3 (CD5)- and R1-10B5 (CD8)-positive cells in methylcholanthrene-induced autochthonous tumor. These CD5- and CD8-positive lymphocytes were also recognized in an N-nitrosourea-induced syngeneic tumor-host system and actually showed specific cytotoxicity against tumor cells in vitro. Macrophages were recognized in tumor tissues more predominantly in the early and terminal phase of tumor rejection; their functions are still uncertain but they are considered to have important immunomodulatory effects. A variety of cytokines were thought to play an important role in augmenting host immunity to achieve tumor rejection. Neutrophils in the tumor tissue were shown to produce a factor attracting lymphocytes to the tumor site, which was designated as lymphocyte migration factor. Subsequently, activities of colony-stimulating factor, interleukin-1, -2, and -3, and cytotoxic-T-cell-generating factor (CGF), which induces final maturation of cytotoxic T cells, were detected at the tumor site as well as in the regional lymph nodes and the spleen. CGF was found to be produced by W3/25 (CD4)-positive T cells. Lymphocytes residing in the spleen of the immune rats did not show cytotoxic activity against tumor cells but significant tumor lysis activity was recognized with TIL. This suggests that lymphocytes may achieve maturation after they leave the spleen. Cellular reactions occurring at the tumor site were enhanced at each step by various cytokines produced by lymphocytes as well as by inflammatory cells. This cytokine cascade seems to be essential for obtaining a sufficient immune response for tumor rejection. When an established T9 subcutaneous tumor with a diameter of 10 mm was treated by intratumoral infusion of lymphokine-activated killer (LAK) cells, the tumor showed complete regression after 2-3 weeks of transient growing.(ABSTRACT TRUNCATED AT 400 WORDS)     

Mast cells of the lungs in experimental hyperergic pleurisy

Reaction of mast cells, the content of free and cell histamine and serotonin in the lung tissue at early stage of inflammation were studied on the model of hyperergic pleurisy in albino rats. Intrapleural antigen injection to sensitized rats was followed by progressive degranulation of mast cells of pleural and subpleural lung tissue with release of histamine and serotonin. The maximal increase in the content of free amines was found after 15 min. The level of free amines did not differ significantly from the initial one by the first hour. The early activation of amines synthesis and their storage recovery were observed as well as reactions of the late phase of immunological activation in the mast cells as a leukocytic tissue infiltration by subsequently polymorphonuclear leukocytes and macrophages.     

POST-MITOTIC AGE OF MONONUCLEAR CELLS MIGRATING INTO TA-3(St) SOLID TUMORS

Subcutaneous transplantation of 5 × 10⁶ TA-3(St) mammary carcinoma cells into A/J mice produced rapidly growing tumors that showed a substantial accumulation of lymphocytes, monocytes and macrophages. This must have resulted primarily from an influx of circulating cells, since none of the cell types exhibited any significant local proliferation as indicated by 1 hr ³H-TdR uptake. The post mitotic age of the various leukocyte types in circulation of normal and tumor bearing animals, and those appearing within the tumors, was evaluated by a repeated ³H-TdR labeling protocol designed to label newly formed leukocytes. Tumor transplantation (or injection of an equivalent volume of saline in control animals) was preceded by thirteen ³H-TdR injections (12.5 μCi every 8 hr) and followed by eight more injections at equivalent intervals. Animals were serially sacrificed at 5-14 days after tumor transplantation or saline injection. Total lymphocyte labeling in the blood during these intervals was higher in tumor bearing mice (approximately 25% between 5 and 12 days) than in the saline injected controls (approximately 20%), indicating that tumor transplantation resulted in an increase in the proportion of young lymphocytes in circulation. More significantly, a much higher labeling (57-60% at 5-12 days) was exhibited by lymphocytes isolated from the tumors, indicating selective migration (and/or retention) of newly formed lymphocytes within the tumor. This finding applied to lymphocytes in all size categories. Although blood monocyte labeling in the control and experimental animals was similar (e.g. 83% at day 5 and 52% at day 14), significantly higher labeling (e.g. 93% and 70% at the respective intervals) was exhibited by monocytes isolated from tumors. This suggested a preferential accumulation of young monocytes at the tumor site. An identical macrophage labeling pattern compared to that shown by monocytes within the tumor indicated a rapid monocyte-macrophage transition. Since these findings in the present strain-specific solid tumor are similar to those previously obtained in this laboratory in the strain-nonspecific Ehrlich ascites tumor, the phenomenon of selective localization of newly formed mono-nuclear leukocytes appears to be a general occurrence in transplanted murine tumors.     

The role of host lymphocytes and host macrophages in antitumor reactions after injection of sensitized lymphocytes and tumor target cells into naive mice

Summary DBA/2 mice were immunized i.p. against syngeneic SL2 lymphosarcoma cells. At various days after the last immunization peritoneal and spleen lymphocytes were collected. The lymphocyte suspensions were enriched for T-cells by nylon wool filtration.The peritoneal T-cells from immunized mice (a) expressed direct specific antitumor cytotoxicity in vitro, (b) induced macrophage cytotoxicity in vitro, and (c) exerted tumor neutralization measured in a Winn-type assay. Spleen T-cells from these immunized mice (a) expressed no direct specific antitumor cytotoxicity in vitro, (b) only induced moderate macrophage cytotoxicity in vitro, but (c) exerted tumor neutralization in a Winn assay.For effective tumor neutralization in vivo effector target cell ratios of 1000:1 were required. When the effector/target ratio of 1000:1 was maintained but the absolute numbers of effector and target cells were lowered from 10⁶ to 10⁵ lymphocytes and 10³ to 10² target cells respectively, no tumor neutralization was obtained.The major effect of the sensitized-transferred T-lymphocytes seemed to be the induction of cytotoxic macrophages in the (naive) recipient mice, as the peritoneal macrophages collected from the recipient mice 7 days after i.p. injection of a mixture of sensitized T-cells and tumor cells were cytotoxic. Purified peritoneal T-lymphocytes collected from these recipient mice were able to induce macrophage cytotoxicity in vitro but expressed no cytotoxic T-cell activity.In conclusion, our results show that in the tumor system used, tumor neutralization after transfer of sensitized lymphocytes is not dependent on the presence of cytotoxic T-lymphocytes. Lymphocytes with the strongest potency to render macrophages cytotoxic (in vitro and in vivo) also induce the best tumor neutralization in vivo, suggesting an important role for host macrophages as antitumor effector cells.     

The multicellular tumor spheroid model: I. Characterization of the allograft response in sensitized mice

Intraperitoneal implants of multicellular spheroids of EMT6 mammary sarcoma cells into specifically sensitized allogeneic mice have been used as a model to study in situ immunity to solid tumor allografts. Morphological examination of the spheroids recovered from sensitized mice revealed massive infiltration by host cells. There was an early but apparently nonspecific infiltration by large numbers of polymorphonuclear leukocytes followed by the infiltration of large numbers of macrophages and smaller numbers of lymphocytes. The later infiltration correlated with tumor cell killing as determined by a cloning assay for surviving EMT6 cells, and with the presence of cells with cytotoxic activity in in vitro assays. The kinetics of tumor cell killing indicated that tumor cell survival decreased within a few hours of spheroid implantation and less than 1% of the tumor cells survived after 48 h. Purification of the infiltrating cells by nylon wool fractionation and by treatment with anti-T-cell serum and complement indicated that the primary cytotoxic cell is a T lymphocyte.     

In situ effector mechanisms in rat kidney allograft rejection. I. Characterization of the host cellular infiltrate in rejecting allograft parenchyma.

The infiltrating inflammatory cells were recovered with collagenase and DNase from rejecting rat kidney allografts and autografts in conditions where the enzyme treatment did not affect the expression of subclass-specific surface markers. As the differential distribution of the inflammatory cells in the dispersate was similar to the distribution of inflammatory cells in tissue imprints, and as any major blood contamination was excluded, we consider the results representative of the composition of the in situ infiltrate. At the peak of rejection on Day 6 after the transplantation, approximately 30% monocytes, 17% macrophages, 31% lymphocytes, 6% (T) lymphoblasts, and 10% (B) plasmablasts and plasma cells were present in the graft. The blast cell response, pathognomonic to immune activation, was less prominent in the recipient spleen, blood, and lymph nodes. Twenty-three percent of the infiltrating lymphocytes expressed the (T-cell-specific) Pta.A.1 surface antigen(s) and 14% were surface Ig positive. The remaining lymphocytes were double-negative “null cells.” In preparative cell electrophoresis most of the allograft-infiltrating lymphocytes carried the low electrophoretic mobility, characteristic to resting B cells. Approximately 70% of allograft-infiltrating macrophages and 50% of infiltrating monocytes but only 30% of the monocytes present in the recipient spleen expressed the Fc receptor to IgG, suggesting an activation (or increase in avidity) of this receptor during the influx of mononuclear cells into the site of inflammation and during maturation of monocytes into tissue macrophages. There was a strong in situ proliferative activity, far stronger than in the central lymphatic system of the recipient rat. After 1 hr in vivo pulse labeling with [3H]thymidine 24% of the infiltrating inflammatory cells carried the label. Most of the labeled cells were blasts or lymphocytes, but a small albeit distinct number of labeled monocytes were also present in situ. In contrast to the recipient spleen, where most of the labeled lymphoid cells had a high electrophoretic mobility of resting T cells, in the infiltrate most of the labeled lymphoid cells had a slow mobility of resting B cells.     

Sensitization of T lymphocytes in vitro by syngeneic macrophages fed with tumor antigens.

We investigated the interaction between T lymphocytes and macrophages in the vitro sensitization of lymphocytes against tumor cells. Spleen cells were sensitized in vitro by syngeneic peritoneal macrophages that had been fed with cell-free antigen preparation of syngeneic tumor cells. The sensitized T lymphocytes acquired specific cytotoxic cells. The sensitized T lymphocytes acquired specific cytotoxic activity in vitro and the capacity to inhingeneic fibroblasts, or the antigen preparation by itself were not able to sensitize the lymphocytes against the tumor.     

Enhancement of human natural killer cell cytotoxicity by serotonin: role of non-T/CD16+ NK cells, accessory monocytes, and 5-HT1A receptors

Serotonin (10(-4) - 10(-7) M) augmented natural killer cell cytotoxicity (NKCC) of human CD16+/non-T lymphocytes in vitro against the NK-sensitive target cells K 562 erythroleukemic, Molt-4 lymphoma, Chang liver cells, and against EBV-transformed Daudi B-lymphoblastoid target cells by a mechanism of action involving a prostaglandin-and IL-1-independent accessory function of monocytes. No evidence for the production of intermediary, NK-enhancing cytokines by serotonin was obtained, suggesting a cell-to-cell-mediated interaction between monocytes and NK cells as a plausible mechanism of action for the NK-augmenting effect. Monocytes recovered by counter-current centrifugal elutriation but not monocytes recovered by adherence reconstituted the effect of serotonin when added to nonadherent NK cells. NK-enhancing effects of serotonin were mimicked by two 5-HT1A-type serotonin receptor agonists, 8-OH-DPAT and (+)-ALK. The development of NKCC in response to serotonin could be resolved into (i) an induction phase, dependent on the presence of accessory monocytes and serotonin, and (ii) an effector phase, independent of the presence of monocytes or serotonin. Serotonin-activated MNC continued to exert augmented cytotoxicity for at least 8 hr after the removal of serotonin and monocytes. In several experiments, serotonin-activated NK cells killed greater than 75% of K 562 target cells even at low effector to target cell ratios and low baseline NKCC. We suggest that serotonin may have a role in nonspecific tumor defence by regulating an earlier unrecognized interplay between monocytes and NK cells.     

Emperipolesis of lymphoid cells by human follicular dendritic cells in vitro.

Isolated follicular dendritic cells (FDCs) showed true and pseudoemperipolesis of fresh tonsillar lymphocytes, even after long-term (50-day) cultivation. Emperipolesis by FDCs was not restricted by allotype specificity, nor was it inhibited by the addition of antibodies against MHC-I & II antigens. Follicular dendritic cells predominantly engulfed B-cells; monocytes and macrophages were not found between FDC cytoplasmic extensions. When highly purified T-cell populations were added to FDC cultures emperipolesis of T-cells occurred, particularly those of the CD4-positive phenotype. Mitoses appeared within 6 h in the emperipolesed lymphocytes and, after an additional 18 h, some lymphocytes exhibited apoptosis.     

Comparison of monocyte and alveolar macrophage antibody-dependent cellular cytotoxicity and Fc-receptor activity

The cytotoxic potential of rabbit peripheral blood monocytes and alveolar macrophages in antibody-dependent cellular cytotoxicity (ADCC) toward both erythrocyte (RBC_ox) and tumor cell (CEM T-lymphoblast) targets was examined. ADCC was measured in a 4-hr ⁵¹Cr-release assay. Alveolar macrophages were more efficient at killing the tumor cell targets (optimally sensitized with rabbit antisera) than monocytes at similar effector cell/target cell ($\text{E}\text{T}$) ratios. Tumor cell targets sensitized with seven different antisera (anti-CEM) were lysed by alveolar macrophages but not by the monocytes. In marked contrast, the monocytes were more effective at lysing the sensitized erythrocyte target cells. The degree of cytolysis of RBCox and CEM was dependent on the $\text{E}\text{T}$ ratio and the degree of sensitization of these target cells. It was demonstrated that the effector cell selectivity in ADCC was directly related to their ability or inability to bind the sensitized target cells as determined by Fc-receptor rosette formation. The transition from monocyte to macrophage may, therefore, have resulted in an alteration in the criteria necessary for Fc-receptor binding to antibody-sensitized target cells and subsequent ADCC.     

Specific T-cell factor production and lymphocytes in the direct surroundings of a subcutaneous allogeneic tumor.

Antigen-specific T-cell factors (TCF) play a role in the initiation of cellular immune responses. In allogeneic mouse-tumor models lymphocytes from the direct tumor surroundings of both euthymic and nude mice produce TCF. These lymphocytes produce TCF when collected already 1 day after subcutaneous (sc) injection of tumor cells. In contrast to euthymic mice, draining lymph nodes and spleen of nude mice did not contain TCF-producing lymphocytes at any stage after sc tumor cell injection. In sensitized euthymic mice TCF production by lymphocytes is significantly higher in the direct tumor surroundings than in draining lymph nodes or spleen. At 2 and 5 days after tumor cell injection, the mononuclear cell infiltrate of the tissue surrounding the tumor in euthymic mice showed low expression of Thy 1, CD3, TCR alpha beta, TCR gamma delta, CD4, CD8, and asialo GM1, whereas several lymphocytes and mast cells were positive for monoclonal antibody (mAb) 14-30 (directed against TCF). In both euthymic and nude mice, sc injected tumor cells showed apoptosis. In conclusion, the direct tumor surroundings are the first (and, for nude mice, the only) site of TCF production, sc injection of tumor cells attracts mAb 14-30-positive lymphocytes and renders mast cells positive for mAb 14-30.     

Separated parabiont reveals the fate and lifespan of peripheral-derived immune cells in normal and ischaemia-induced injured kidneys

Immune cell infiltration plays a key role in acute kidney injury (AKI) to chronic kidney disease (CKD) progression. T lymphocytes, neutrophils, monocytes/macrophages and other immune cells regulate inflammation, tissue remodelling and repair. To determine the kinetics of accumulation of various immune cell populations, we established an animal model combining parabiosis and separation surgery to explore the fate and lifespan of peripheral leucocytes that migrate to the kidney. We found that peripheral T lymphocytes could survive for a long time (more than 14 days), whereas peripheral neutrophils survived for a short time in both healthy and ischaemia-induced damaged kidneys. Nearly half of the peripheral-derived macrophages disappeared after 14 days in normal kidneys, while their existing time in the inflammatory kidneys was prolonged. A fraction of F4/80^high macrophages were renewed from the circulating monocyte pool. In addition, we found that after renal ischaemia reperfusion, neutrophils increased significantly in the early phase, and T lymphocytes mainly accumulated in the late stage, whereas macrophages infiltrated throughout AKI-CKD progression and were sustained longer in injured as opposed to normal kidneys. In conclusion, peripheral-derived macrophages, T lymphocytes and neutrophils exhibit different lifespans in the kidney, which may play different roles during AKI-CKD progression.     

Macrophage-mediated in vitro sensitization of lymphocytes

Summary Antigen-fed macrophages were able to induce specific sensitization of unprimed syngeneic lymphocytes in vitro. The sensitized lymphocytes caused specific injury to target cells that carried the relevant antigens. In the present study, we investigated the in vivo activity of lymphocytes sensitized by antigen-fed macrophages. Mouse spleen cells were sensitized by macrophages that had been exposed to the radiation leukemia virus (RadLV). The sensitized lymphocytes, which were enriched for T-cells, were injected to syngeneic normal recipients and 4 days later the mice were challenged with RadLV-induced lymphoma cells. By following tumor growth and survival of mice, we have found that the sensitized lymphocytes protected the recipient mice against lymphoma development if injected 4 days before the tumor cells. The protective activity of the sensitized lymphocytes was radioresistant, but they could not protect irradiated hosts. It is suggested that macrophagemediated in vitro sensitization of lymphocytes induces initiator cells that can protect the recipient host by recruitment of a defensive immune response.