Combination chemo-immunotherapy: kinetics of in vivo and in vitro generation of natural killer cells and lymphokine-activated killer cells in the rat.

Rats received a single high dose of cyclophosphamide (Cy) (150 mg/kg), followed 48 h later (on day 0) by immunization with a T cell-dependent soluble antigen, ovalbumin in Freund's complete adjuvant (FCA). The effect of this treatment on lymphoid cell subpopulations in the spleen, natural killer (NK) cell and interleukin-2 (IL-2) induced lymphokine-activated killer (LAK) cell activity was examined. Cy (with and without ovalbumin) caused a large relative increase (by day 14) in splenic OX8+, OX19- cells with NK morphology. A marked relative increase in fresh NK cell activity was noted after Cy + ovalbumin, but not consistently after Cy alone. Elevated NK activity was Cy dose- and time-dependent, was evident within 7 days post Cy/ovalbumin and persisted for at least 28 days. Pooled splenic mononuclear cells (MNC), obtained 14 days after Cy/ovalbumin, lost all cytolytic activity against YAC-1 cells when cultured in the absence of human recombinant IL-2 (rIL-2). In contrast, similarly maintained cells from normal rats displayed NK activity higher than normal 'fresh' levels. Upon culture in medium containing 500 U/ml rIL-2, however, 'augmented' NK activity was equivalent, on a per-cell basis, in both normal and Cy/ovalbumin-pretreated groups. LAK activity generated in vitro (i.e. against NK-resistant target cells) was significantly lower in the latter group, and the overall yield of cells was reduced. By day 21 after Cy/ovalbumin, augmented NK activity was significantly greater than controls, on a per-cell and total culture yield basis. Moreover, LAK activity was now similar between groups. It is concluded that the chemotherapy/immunization protocol which we have used can greatly enhance NK activity in vivo and that these cells are responsive to induction of LAK activity by IL-2 in vitro.     

The role of natural killer cells in resistance to coccidiosis: investigations in a murine model.

Natural killer (NK) activity, detected by the lysis of Yac-1 target cells, was examined in splenic and mesenteric lymph node (MLN) cells throughout the course of infection with Eimeria vermiformis in BALB/c and C57B1/6 (B6) mice. These strains are, respectively, relatively resistant and susceptible to primary infections, which render them equally, and completely, resistant to challenge. Resting levels of NK activity were higher in B6 than in BALB/c, and B6 responded earlier in the course of infection than BALB/c, but splenic peak values were higher in BALB/c; the pattern of response in MLN cells was similar in both strains, but the peak was higher in BALB/c. At the time (7 days p.i.) of peak NK response in BALB/c mice there was, depending upon the choice of NK-resistant/lymphokine-activated killer (LAK)-sensitive target cells, either little (P388D1), or no (P815) splenic LAK activity. Challenge of immunized BALB/c mice did not evoke a detectable NK response. Although the higher NK activity in BALB/c mice correlated with greater control of primary infection, depletion of NK activity (demonstrated in splenic cells) in vivo by treatment with anti-asialo GM1 antibodies did not greatly affect the course of infection. Furthermore, this treatment did not augment the exacerbation of infection produced by treatment with anti-interferon-gamma (IFN-gamma) MoAb, indicating that, at least in this system, NK cells are not a fundamentally important source of this controlling cytokine of eimerian infections. The results suggest that NK cells may not greatly influence the outcome of coccidial infections.     

Cure of human melanoma lung metastases in nude mice with chronic indomethacin therapy combined with multiple rounds of IL-2: characteristics of killer cells generated in situ

We have shown that macrophage-derived prostaglandin (PG)E2 inactivates all interleukin 2 (IL-2) dependent killer cell lineages in the tumor-bearing host, so that chronic indomethacin therapy (CIT) combined with multiple rounds of IL-2 can cure experimental and spontaneous metastases of a variety of murine tumors. We tested the efficacy of this therapy on experimental human melanoma metastasis in nude mice and characterized the killer cells generated in situ. BALB/c nude mice were injected i.v. with 2 x 10(6) human lung-metastasizing line P52, MeWo melanoma cells. After 5 weeks, when lung nodules were well established, mice received vehicles alone (control) or were given (a) CIT (14 micrograms/ml in drinking water); (b) three rounds of IL-2, 25,000 Cetus U, 8 hourly i.p. (days 40-44, 50-54, 60-64); (c) CIT + three rounds of IL-2; (d) CIT + four rounds of IL-2 (round 4 on days 70-74); and (e) CIT + five rounds of IL-2 (round 5 on days 80-84). Control and experimental mice were killed on day 71 to score lung colonies and evaluate killer activity in splenic and lung lymphocytes and macrophages against murine YAC-1 lymphoma and B16F10 melanoma, human P52 melanoma, K562 erythroleukemia, and Raji lymphoma targets. Killer cells for P52 were phenotyped for Thy-1, Lyt-2, and asialo-GM-1 markers by ab + C'-mediated deletion of killer function. Mice in all groups were also kept for survival. CIT alone improved splenic NK activity but marginally reduced the lung colony counts or prolonged the survival time. Three rounds of IL-2 alone reduced the median colony counts by 50% and prolonged the survival by 2 weeks, but resulted in no long-term, disease-free survival, in spite of significant activation of LAK cells with Thy-1-, Lyt-2-, AGM-1+ phenotype in the spleen. CIT + 3 rounds of IL-2 reduced the median colony counts from 40 to 0 and improved the survival from a median of 66 (control) to 120 days (40% surviving 260 + days). CIT + four or five rounds of IL-2 caused long-term (260 + days) survival of 80% mice, most surviving 400 + days. The combination therapy activated killer lymphocytes (Thy-1-, Lyt-2-, AGM-1+) and, to a smaller extent, macrophages (AGM-1 +/-) in the spleen and the lungs, showing a high cytocidal ability for all the targets.(ABSTRACT TRUNCATED AT 400 WORDS)     

rIL-4 differentially regulates rIL-2-induced murine NK and LAK killing in CD8+ and CD8- precursor cell subsets

Interleukin 4 (IL-4) and IL-2 have complementary or synergistic roles in many aspects of lymphocyte development. IL-2 supports the induction of cytolytic activity in cytotoxic T lymphocyte (CTL), natural killer (NK), and lymphokine-activated killer (LAK) cells. IL-4 has also been shown to support CTL and LAK in primary murine spleen cell culture. This report demonstrates that IL-4 selectively down-regulates IL-2 inducible murine CD8- precursors of NK cells. For maximal regulatory effect it is necessary to add IL-4 to cultures before 40 h. Enrichment for NK1.1+ cells failed to recover precursor cells which are down-regulated in overnight cultures or can be cultivated in vitro to yield NK cytolytic activity. Furthermore, phenotypic analysis of effector cells demonstrated a marked inhibition of development of NK1.1+ cells in cultures containing IL-4 plus IL-2 versus IL-2 alone. Thus, it appears that IL-4 down-regulates the precursors of murine NK cells by inhibiting proliferation and/or development. In addition, we show that IL-2-induced murine LAK activity mediated by CD8- precursor cells is unaffected by IL-4, while CD8(+)-derived LAK cells are up-regulated by co-culture with IL-4 and IL-2. Analysis of these data relative to reports documenting down-regulation of human LAK by IL-4 suggests that in vitro cultured, IL-2-activated murine NK cells are the correlates to what are commonly described as human LAK cells. The discrepancy may stem from differences in the characteristics of target cells used in the murine versus the human systems. These results clarify the conflicting reports on the effect of IL-4 on killing activity.     

Lymphokine-activated killer (LAK) cells modulate the effects of IL-2 on a T cell-mediated immune response.

The ability of LAK cells and/or IL-2 to affect the course of an established T cell response was examined in a delayed-type hypersensitivity (DTH) model. IL-2 greatly increased the magnitude of the response at 24 h, while LAK cells alone had no effect. The administration of LAK cells and IL-2 together also had no effect on the magnitude of the DTH response, demonstrating that LAK cells were able to remove the enhancement seen with IL-2 alone. The presence of LAK cells reduced the serum half-life of IL-2 significantly, but not to an extent able to account for the observed loss of IL-2 induced DTH enhancement. IL-2 administration influenced cell phenotypes in the spleen and draining lymph nodes (DLN), as well as increasing splenic weight; the additional presence of LAK cells markedly altered these effects of IL-2 in the spleen (but not the DLN). Taken together, these results suggest that LAK cells interact with activated T-cells within the immune system and modulate their function.     

Natural killer and interleukin-2 induced cytotoxicity in asthmatics

Natural killer (NK) cell activity and interleukin-2 (IL-2) induced cytotoxicity to NK-resistant targets T24 in a group of patients with grass pollen seasonal asthma were investigated. Acute bronchial provocation with this antigen was also performed and blood samples were taken before and 15 min after the maximal clinical response. NK activity was elevated in this group but there was a significant decrease after acute challenge. IL-2 induced cytotoxicity was also increased and declined after antigen challenge. Limiting dilution analysis for minimal frequencies of precursors of the IL-2 induced killer cells was introduced and showed increased numbers of lymphokine-activated killing precursors in this group of patients. These data might indicate the aberrations of the IL-2 system in immediate type allergy but the clinical significance of these findings remains to be determined.     

Signal transduction via phosphorylated adhesion molecule, LFA-1{beta} (CD18), is increased by culture of natural killer cells with IL-2 in the generation of lymphokine-activated killer cells

It is well known that IL-2 stimulates natural killer (NK) cellsto express lymphokine activated killer (LAK) activity and thatthis stimulation prompts the acquisition of the ability to lysepreviously insensitive target cells. The possible role of adhesionmolecules in the IL-2 activation process was probed by focussingon a lymphocyte function-associated antigen (LFA)-1-dependentmodel system. A mAb to the LFA-1ß chain abrogatedLAK activity, but only moderately suppressed NK activity, suggestinga differential role for LFA-1ß In LAK compared withNK mediated lysis. Orthophosphate labeling demonstrated thatthe LFA-1ß chain was strongly phosphorylated in LAKbut not NK cells; in contrast, the chain was phosphorylatedsimilarlyin both effector cell types. At least a portion ofthe phosphorylation of the ß chain was on tyrosineresidues, as shown by Western blotting with anti-phosphotyrosineantibody of LFA-1ß immunoprecipitates. Crosslinkingof the LFA-1ß chain with plastic-adhered antibodystimulated Ca²⁺-dependent release of cytoplasmic lytic granulesand induced phosphatidyl inositol turnover in LAK but not NKcells. We conclude that the IL-2-induced phosphorylation oftheß chain of the LFA-1 adhesion molecule in LAK cellsand associated alteration in signal transduction may be importantin the stimulation of LAK cell activity in NK cells.     

Age-associated decline in IL-2 and IL-12 induction of LAK cell activity of human PBMC samples

Previously we reported that young and elderly natural killer (NK) cell activity against the standard NK sensitive K562 cell line can be augmented to the same degree by IL-2 and IFN-. We have extended these studies to include IL-12. Similar to IL-2 and IFN-, IL-12 can enhance NK cytotoxicity to the same degree in both young and elderly samples over a wide range of doses and incubation times when K562 cells are used as targets. However, in contrast to our findings with the NK system, we have observed that induction of lymphokine activated killer (LAK) cell activity, as defined by the ability of peripheral blood mononuclear cells (PBMC) samples to lyse the normally NK resistant Daudi cell line, was significantly decreased in the elderly samples compared to young samples. Comparable age-associated differences were observed in LAK activity after induction with IL-2, IL-12, and IFN- at varying doses and incubation times. We hypothesize an age-associated deficiency either in the mechanism of LAK induction or in target cell recognition.     

Human pulmonary natural killer (NK) cells exhibit limited lymphokine-activated killer (LAK) activity

The spontaneous activity of natural killer (NK) cells against most solid tumor targets is low but can be increased by incubation with interleukin 2 (IL-2). This phenomenon, termed lymphokine-activated killer (LAK) activity, has been used in recent clinical trials against some pulmonary malignancies. We compared the LAK activity of blood and lung lymphocytes after activation with IL-2. Lung lymphocytes did not develop LAK activity despite demonstrating a significant increase in NK activity against K562 targets after incubation with IL-2. This functional difference correlated with a reduced expression of Leu-19, a marker present on virtually all LAK cells derived from peripheral blood, on lung NK cells. Because pulmonary macrophages (PM) are important regulators of NK function, we next investigated whether PM could be responsible for the functional and phenotypic differences noted. Measuring NK and LAK activity in parallel, we found that the addition of PM to IL-2-activated lymphocytes resulted in a preferential suppression of LAK activity and a loss of Leu-19 expression from IL-2-activated blood lymphocytes as well as a Leu-19+ T cell clone. We conclude that pulmonary NK cells are phenotypically and functionally different from peripheral blood NK cells and that this likely reflects local regulation, perhaps by PM.     

Effects of in vitro treatment with fluticasone propionate on natural killer and lymphokine-induced killer activity in asthmatic and healthy individuals

BACKGROUND: Topical corticosteroids are beneficial in the treatment of allergic respiratory disorders; they exert effects on a number of cells involved in allergic inflammatory reactions. On the other hand, major histocompatibility complex (MHC)-unrestricted cytotoxicity (i.e., natural killer [NK] cell activity) may play a role in the inflammatory allergic reaction. The objective was to gain insight into the mechanisms of the therapeutic effects of fluticasone propionate (FP), an inhaled corticosteroid used in asthma and rhinitis therapy. Therefore, we evaluated the NK and lymphokine-activated killer (LAK) activity of effector cells in vitro treated or not with FP. METHODS: Evaluations were made on peripheral blood mononuclear cells (PBMNCs), obtained from healthy volunteers (n = 10) and from asthmatic atopic subjects (n = 10) with allergy to Parietaria. RESULTS: Asthmatic patients had significantly increased NK activity (P= 0.0008), and interleukin (IL)-2- (P=0.0005) and interferon (IFN)-alpha-induced LAK activities (P=0.0005). In both groups, FP 10(-7) M significantly reduced NK activity (P<0.0001), IL-2-induced LAK activity (P<0.0001), and IFN-alpha-induced LAK activity (P<0.0001). Similar results were obtained with FP 10(-8) M. CONCLUSIONS: Since MHC-unrestricted cytotoxicity has been implicated in the development of allergen-induced eosinophilic airway inflammation, inhibition of NK and LAK activity by FP may contribute to the steroid therapeutic effect in asthma.     

Protein A potentiates lymphokine-activated killer cell induction in normal and melanoma patient lymphocytes.

The effects of purified protein A from Staphylococcus aureus Cowan I stain on induction of lymphokine (IL-2) activated killer (LAK) activity were studied in normal as well as melanoma patient's lymphocyte. The coculture of peripheral blood mononuclear cells (PBMC) with various doses of protein A (0.001, 0.01 and 0.1 microgram/ml) and IL-2 (100 U/ml) for 4 days produced synergistic effect on the LAK cells mediated cytotoxicity. The potentiation of cytotoxicity and lytic ability of LAK cells against NK sensitive (K-562) and NK-resistant (M14) tumor cells were observed. Further there was potentiation of DNA synthesis in PBMC after 4 days culture. Similar results were found when PBMC from melanoma patients were cultured with PA and IL-2. The potentiation of LAK cell induction associated with its cytotoxic and lytic potential by low doses of IL-2/PA regiment may be helpful in the development of LAK immunotherapy of the cancer patients.     

Peripheral blood lymphocytes from thermal injury patients are defective in their ability to generate lymphokine-activated killer (LAK) cell activity

We have previously shown that natural killer (NK) cell activity against K562 tumor cells is severely depressed in thermal injury patients. In this study we have investigated whether the low NK cell activity present in peripheral blood lymphocytes (PBL) from thermal injury patients could be enhanced byin vitro culture with interleukin 2 (IL2) and whether PBL obtained from these patients could generate lymphokine-activated killer (LAK) cell activity against NK insensitive tumor targets. NK cell activity in PBL obtained from 12 different patients was greatly enhanced against K562 tumor cells afterin vitro culture with IL2 for 3 days. In contrast, PBL obtained from these patients and incubated with IL2 had little to no cytotoxic activity when measured against a number of NK-insensitive tumor targets. The failure of PBL obtained from thermal injury patients to generate LAK cell activity was observed regardless of the culture time or the amount of IL2 added to the cultures. PBL from thermal injury patients demonstrated reduced proliferative responses to IL2 and, more importantly, contained suppressor cells which could inhibit the generation of LAK cell activity of normal PBL obtained from control individuals. These results clearly show that in some thermal injury patients NK cell activity can be enhanced by IL2 but these patients are defective in their ability to generate LAK cell activity.     

Interleukin-2 induced killer cell activity against Epstein-Barr virus-immortalized human B cells

Interleukin-2 (IL-2) activated killer (LAK) cells, generated in vitro by treating peripheral blood lymphocytes (PBL) with human IL-2, are able to lyse a wide variety of target cells without restriction by major histocompatibility complex (MHC) molecules. Earlier observations from this and other laboratories indicated that patients with Epstein-Barr virus (EBV) induced infectious mononucleosis, a self-limiting viral disease, have high EBV-nonspecific natural killer (NK) cell activity. Since the effect of LAK cells on EBV-immortalized B lymphocytes has not yet been studied, we decided to investigate LAK cell activity against autologous and heterologous B lymphocytes immortralized in vitro by EBV and other EBV genome-positive and -negative targets of malignant origin. LAK activity was determined by ⁵¹Chromium release assay. The results obtained show that LAK activity was not specific for EBV and was not MHC-restricted. Results of experiments using NK cell reactive monoclonal antibodies suggest that the cytotoxicity is due predominantly to activated NK cells. Our observations suggest that LAK cells may be very effective for immunotherapy in patients with chronic or progressive EBV infections and EBV-induced lymphoproliferative diseases.     

Prostaglandin E2-mediated inactivation of various killer lineage cells by tumor-bearing host macrophages

We have previously reported that natural killer (NK) lineage cells are progressively inactivated during tumor development by prostaglandin E2 (PGE2) secreted by host macrophages; that this facilitates spontaneous tumor metastases, which can be prevented by chronic indomethacin therapy (CIT); and that CIT combined with multiple rounds of interleukin 2 (IL-2) can cure experimental metastases and activate all killer lineage cells in situ including NK cells, lymphokine-activated killer (LAK) cells, and tumoricidal macrophages. The present study tested whether PGE2 secreted by tumor-bearing host macrophages exerts pansuppressor effects against the activation of T cells, NK cells, LAK cells, and tumoricidal macrophages from normal splenic cell populations. Macrophages isolated from CBA mice bearing 21-day intraperitoneal Ehrlich ascites tumors (EAT) or C3H/HeJ mice bearing 21-day subcutaneous T58 mammary adenocarcinomas were added (+/- 10(-5) M indomethacin, or a monoclonal anti-PGE2 ab) to syngeneic splenic lymphocytes to examine the effects on 1) polyclonal activation (3-d 3H-thymidine [3H-TdR] uptake) with concanavalin A (Con A); 2) one-way (CBA alpha BALB/C or C3H alpha BALBC) MLR (5-d 3H-TdR uptake) and subsequent CTL generation (tested against 51Cr-labeled Con A blasts of the stimulator phenotype); 3) NK activity (after 24-h co-culture) against YAC-1 targets; 4) generation of LAK cell activity (in the presence of 200 or 2,000 units recombinant IL-2 for 3 or 5 days), tested against NK-sensitive and NK-resistant targets. Similar effects were also noted on the generation of tumoricidal activity in normal splenic macrophages cultured for 3 days in the presence of LPS. Normal splenic macrophages added under the same conditions served as controls. Effects of pure PGE2 or PGF2 alpha (10(-6) M) were also examined on these activation events. Results revealed that tumor-host-derived macrophages (but not normal macrophages) markedly suppressed all these activation events and this suppression was abrogated nearly totally by indomethacin and totally by anti-PGE2 ab, indicating its mediation by PGE2. This finding ran parallel with high levels of PGE2 production by tumor-host-derived but not normal splenic macrophages. Pure PGE2 but not PGF2 alpha mimicked these suppressor effects. While tumoricidal activity was generated in normal macrophages in the presence of LPS, IL-2, or IFN-gamma or their various combinations (which led to further augmentation), these agents required the presence of indomethacin to generate significant killer activity in tumor-host-derived macrophages. macrophages.(ABSTRACT TRUNCATED AT 400 WORDS)     

Dissociation of natural killer and lymphocyte-activated killer cell lytic activities in human CD3− large granular lymphocytes

CD3^? large granular lymphocytes (LGL) are known to display natural killer cell (NK) activity without prior sensitization or restriction by major histocompatibility antigens. Upon short-term exposure to interleukin-2, NK cells were shown to acquire lymphocyte-activated killer cell (LAK) activity. The aim of this study was to analyze the characteristics of these lytic activities. Our data indicated that both NK and LAK activities were Ca²⁺ dependent; however, they could be dissociated by a Ca²⁺ channel blocker or a Ca²⁺ channel competitor agent. Moreover, NK activity was associated with granule exocytosis of lytic proteins spontaneously present in CD3^? LGL, the most likely candidate being the pore-forming protein perforin. By contrast, LAK activity was found to be dependent on de novo protein synthesis and distinct from granule exocytosis. Our results strongly suggest that NK and LAK activities could be defined as two distinct pathways involving different lytic mediators.     

Interleukin-2 modulates the expression of lymphocyte function-associated antigen-one (LFA-1) and p150,95 during the generation of lymphokine-activated killer (LAK) cells

Lymphocyte function-associated antigen-one (LFA-1), Mac1 and p150,95 represent a family of heterodimeric cell surface molecules with a common beta subunit and distinct alpha subunits. LFA-1 is known to be functionally important in cell-cell interactions between immune cells. In the present study, a mouse monoclonal antibody (mAb), RH1-38, which recognizes an epitope on the beta-chain of LFA-1 was used to study the function and expression of LFA-1 on lymphokine-activated killer (LAK) cells. This mAb has been shown previously to block, in the absence of complement, cytolytic activity mediated by natural killer (NK) cells, cytotoxic T lymphocytes (CTL), and a monocyte-like cell (phorbol diester-stimulated HL-60 cells). LAK cells were generated by culturing in vitro human peripheral blood lymphocytes (PBL) in the presence of human recombinant interleukin-2 (rIL-2), and cytotoxic activity was measured by a 51Cr-release assay using the human NK-resistant Daudi cell line. Addition of RH1-38 ascites supernatant, purified RH1-38 mAb, or F(ab')2 fragment of RH1-38 markedly reduced (>80) LAK cytolytic activity, whereas NS-1 (parent hybridoma) ascites supernatant, normal mouse IgG, and monoclonal anti-HLA had no effect on LAK-mediated killing. Equivalent inhibition of NK and CTL activity by purified RH1-38 required 10-100-fold more antibody. Appreciable inhibition occurred if the mAb was added up to 2 hr after LAK cells were mixed with targets. Indirect immunofluorescence flow cytometry and immunoprecipitation studies revealed that LFA-1 and p150,95 expression were dramatically enhanced in PBL populations cultured with rIL-2 compared with PBL cultured without rIL-2; Daudi cells expressed no detectable LFA-1 family heterodimers. Time-course experiments demonstrated that during culture of PBL in the presence of rIL-2, development of enhanced expression of LFA-1 and p150,95 correlated closely with LAK cytolytic activity. These studies (i) demonstrate that LFA-1 and/or p150,95 are functionally important effector cell surface molecules expressed by LAK cells and that some homology to NK and CTL mechanisms of cell-mediated lysis may exist; and (ii) suggest that enhanced LFA-1 and/or p150,95 expression are important for development of the fully differentiated LAK effector cell in the presence of rIL-2.     

慢性乙肝患者杀伤性免疫细胞功能的研究

通过对４４例病毒性肝炎患者Ｔ细胞亚群，ＮＫ细胞活性与ＬＡＫ细胞活性的观察，探讨了在慢性乙型肝炎病毒复制与非复制状态下的杀伤性细胞活性。结果表明：在乙肝病毒的高复制状态下，ＣＤ８＾＋细胞数增加，ＣＤ４＾＋／ＣＤ８＾＋比例显著下降；ＮＫ细胞活性与ＬＡＫ细胞活性也明显低下，且在ＨＢｅＡｇ与ＨＢＶＤＮＡ阳性组中，ＮＫ活性与ＬＡＫ活性的改变与ＨＢｅＡｇ的Ｐ／Ｎ值变化呈显著负相关，而ＮＫ活性与ＬＡＫ活性变化则     

Lymphokine-activated killer (LAK) cell activity in tumor-infiltrating lymphocytes from non-small cell lung cancer

Tumor-infiltrating lymphocytes (TILs) are often seen in non-small cell lung cancers (NSCCs). Their functional role in the pathogenesis of lung cancer is unknown. The authors studied TILs in 27 patients with NSCC and determined the following: (1) the immunologic phenotype as defined by monoclonal antibodies against various surface markers, (2) activation state as indicated by interleukin-2 (IL-2) receptor expression and the kinetics of proliferation response to IL-2, and (3) the ability to develop lymphokine-activated killer (LAK) type cytotoxicity against both natural killer (NK)-resistant tumor cell targets (M14) and fresh autologous tumor cells. The authors' results show TILs from NSCCs to be a heterogeneous population composed of T-cells, B-cells, monocytes, and NK cells in frequencies similar to those found in peripheral blood lymphocytes (PBLs). TILs demonstrated increased IL-2 receptor expression and a more rapid proliferative response to IL-2 than PBLs, implying activation of TILs by the tumor milieu. Finally, TILs generated cytotoxicity against NK-sensitive (K562) and NK-resistant (M14) cell line targets consistently after in vitro treatment with IL-2 but were less consistent in their ability to lyse fresh autologous tumor cells and less effective than PBL LAK cells in lysing all targets. Comparison with LAK cells generated from normal volunteers suggests that decreased killing of autologous tumor cells only partially results from an inherent resistance to lysis by fresh NSCC targets. It appears, therefore, that tumor cells taken from NSCCs are not readily killed by the immune cells that infiltrate the tumor stroma and that this failure does not result from nonspecific immune deficiency in TILs.     

rhIL-15和rhIL-2诱生的LAK细胞特性比较

目的：研究ｒｈＩＬ－１５激活的ＬＡＫ细胞的增殖、细胞毒作用及相关表型的变化。方法：通过用不同浓度的ｒｈＩＬ－１５和ｒｈＩＬ－２分别诱生 ＬＡＫ细胞，研究二者诱生的 ＬＡＫ细胞的增殖状况；当 ｒｈＩＬ－２和 ｒｈＩＬ－１５为 １５００ Ｕ／ｍｌ时，ＭＴＴ法检测 ＬＡＫ细胞的细胞毒作用，利用流式细胞仪检测细胞表面ＣＤ分子表达情况。结果：ＬＡＫ细胞的增殖对ｒｈＩＬ－２和ｒｈＩＬ－１５均具有时间和剂量依赖性，且无明显差异；终浓度为 １５００ Ｕ／ｍｌ时，ｒｈＩＬ－１５诱生的 ＬＡＫ细胞杀伤 Ｋ５６２细胞的能力强于 ｒｈＩＬ－２诱生的 ＬＡＫ细胞，而对Ｒａｊｉ细胞的杀伤活性，ｒｈＩＬ－２诱生的ＬＡＫ细胞却明显高于 ｒｈＩＬ－１５诱生的 ＩＡＫ细胞，且二者杀伤活性均具有时间依赖性；同时ｒｈＩＬ－１５诱生的ＬＡＫ细胞的ＣＤ３－ＣＤ５６＋细胞、Ｃｄ９４＋细胞百分率均高于ｒｈＩＬ－２诱生的ＬＡＫ细胞。结论：ｒｈＩＬ－１５在体内对ＮＫ细胞功能可能具有较强的调节作用。