Biomedical subjects

T J Eberlein

Publications and source records attributed to T J Eberlein.

At least 91 records · Page 5Linked to original sources

PubMed1991

The role of interleukin-2 in cancer immunotherapy.

The arena of cellular immunotherapy is an emerging field of study. The field has strong foundations in numerous animal tumor models, which provide avenues of research for refinements in human immunotherapy. Experience from a number of research centers can be generalized: LAK cell trials in humans have demonstrated that PBMCs activated with IL-2 possess reproducible antitumor activity in vitro. Exogenous IL-2 administration to patients is required to maintain the biologic activity of the activated cells, although the requirement for LAK cells in tumors like melanoma is not clear-cut. Overall, the response rate to immunotherapy in humans is approximately 30%. The toxicity of immunotherapy, which is related to the dose of systemic IL-2, can be significantly reduced. The next generation of trials of cellular immunotherapy in humans will use TILs. Murine experiments indicate that this population of cells is more powerful and potentially more specific for tumor than LAK cells. Like LAK cells, TILs also require exogenous IL-2 for optimum performance in vivo. The difficulty in reproducible TIL growth can be overcome by the use of monoclonal antibody activation and IL-2. Improvements in TIL therapy are anticipated from the use of TILs combined with hybrid antibodies, the addition of other biologic response modifiers, and the implementation of genetically engineered cells and cell products.

Animals↗

PubMed1990

Biopsy of occult breast lesions. Analysis of 1261 abnormalities.

From January 1, 1987, to December 31, 1988, roentgenographically guided biopsies of 1261 occult breast abnormalities were performed. There were 626 (49.6%) masses and 635 (50.4%) lesions with calcification as the dominant feature. Of the total, 237 (18.8%) were found to be malignant, including 85 cases of ductal carcinoma in situ. Biopsies performed for a mass yielded a 20.8% (130/626) incidence of cancer; and for calcification, 16.9% (107/635). The biopsy results were supported by the interpretations of four radiologists (J.E.M., P.C.S., and M.R.S.) who were practicing at Brigham and Women's Hospital in 22.2% of the total (280), whereas the remaining 981 lesions were discovered on mammograms that were performed and interpreted elsewhere and were referred to our hospital for surgical consultation. The biopsy results of the mammograms from other facilities yielded a positive predictive value of 16.7% (164/981) vs 26.1% (73/280) for the Brigham and Women's Hospital group. This statistically significant difference lends support to the value of a second opinion in patients with biopsy recommendations for occult breast lesions, especially when findings are inconclusive.

Boston↗

PubMed1990

Solid-phase anti-CD3 antibody activation of murine tumor-infiltrating lymphocytes.

Seventeen consecutive s.c. murine tumors, derived from a sarcoma and a colon adenocarcinoma, were cultured in the presence of recombinant interleukin 2 (rIL-2) for growth of tumor-infiltrating lymphocytes (TIL). Identical cultures were activated by solid-phase monoclonal antibody directed against the murine CD3 epsilon-chain, in conjunction with rIL-2. Forty-eight h later, cells were replaced in rIL-2 alone. Proliferation of anti-CD3-stimulated cultures was 1- to 17-fold greater than those cultured with rIL-2 alone (P less than 0.05). Both culture conditions yielded TIL which stained greater than 80% Thy-1.2+/Lyt-2+ (P greater than 0.05), less than 7% Thy-1.2+/L3T4+ (P greater than 0.05). Regardless of culture condition, longitudinal studies of in vitro cytotoxicity generated from 10 TIL preparations revealed no significant differences between the ability of TIL to lyse the murine natural killer-sensitive line YAC or heterologous or autologous tumor (P greater than 0.05). In vivo antitumor activity of TIL was tested by the adoptive transfer of suboptimal doses of TIL plus systemic rIL-2 to mice with pulmonary micrometastatic disease. No difference in tumor regression was noted between the TIL cultured with anti-CD3 plus rIL-2 or with rIL-2 alone (P greater than 0.05). Anti-CD3 stimulation of murine TIL cultures significantly increases lymphocyte cell yield without alteration of their phenotype, in vitro tumoricidal activity, or in vivo therapeutic effect.

Animals↗

PubMed1990

Activation of human tumor-infiltrating lymphocytes by monoclonal antibodies directed to the CD3 complex.

We have used high concentrations of recombinant-methionyl human interleukin 2 (rIL-2) for the initial growth and expansion of human tumor-infiltrating lymphocytes (TIL). Early in the life of the TIL bulk culture, cytotoxicity was non-major histocompatibility complex restricted. Under these culture conditions antitumor cytotoxicity was observed to decline with increasing age of the bulk culture. In addition, TIL became refractory to rIL-2-induced expansion. We have used solid-phase anti-CD3 antibodies for TIL activation followed by culture in reduced concentrations of rIL-2 to reactivate TIL previously grown in high concentrations of rIL-2. TIL refractory to rIL-2 in terms of growth and antitumor cytotoxicity proved sensitive to anti-CD3 activation. The use of solid-phase anti-CD3 was also more effective than high concentrations of rIL-2 in the expansion of TIL when used at the start of culture. Finally, TIL could be induced to secrete IL-2 following solid-phase activation with anti-CD3. These data suggest that human TIL are susceptible to activation by signals directed at the CD3 complex of the TIL cell surface.

Antibodies, Monoclonal↗

PubMed1990

Predictors of local recurrence following conservative breast surgery and radiation therapy. The influence of tumor size.

Size of tumor has not been established as a predictor of tumor recurrence in the breast following conservative surgery and radiation therapy. We analyzed 783 patients with infiltrating carcinoma treated with simple excision and radiation therapy. Median follow-up was 91 months. Median age at diagnosis was 50 years. There was a 13% recurrence among patients with T1 lesions compared with a 12% recurrence among patients with T2 tumors. Size did not predict for local recurrence when the tumor was analyzed by 1-cm increments and whether the tumor was estrogen receptor protein positive or estrogen receptor protein negative. Patients with an extensive intraductal component had a significantly higher local recurrence rate for every tumor size compared with patients with extensive intraductal component-negative tumors. We concluded that size did not predict local recurrence.

Breast Neoplasms↗

PubMed1989

Human tumor-infiltrating lymphocyte (TIL) cytotoxicity facilitated by anti-T-cell receptor antibody.

Long-term growth of tumor-infiltrating lymphocytes (TIL) in high concentrations of rIL-2 is required for generation of therapeutic numbers of cells for adoptive immunotherapy of human cancer. Under these conditions rIL-2 promotes both anti-tumor cytotoxicity and lymphocyte growth from tumors of several histological types. In a series of 16 consecutive tumors, studies of TIL-mediated cytotoxicity against different tumor targets were characterized by an initial strong tumor-non-specific cytotoxicity. With time, TIL bulk cultures became non-cytotoxic against all targets (median time = 38 days). Non-cytotoxic TIL bulk populations were capable of mediating strong cytotoxic responses if pre-treated with anti-T-cell antigen receptor antibody (TcR) before addition to targets. TIL populations were not, however, uniformly susceptible to anti-TcR-mediated cytotoxicity. Anti-TcR-mediated cytotoxicity was confined to CD8+ bulk populations (defined as populations with a CD4/CD8 ratio less than 1), with virtually no cytotoxicity observed in CD4+ populations (CD4/CD8 ratio greater than 2, (p less than 0.01). Both CD4 and CD8 populations expressed TcR antigen reactive with anti-TcR antibody. These results indicate that, despite poor in vitro anti-tumor cytotoxicity in short-term assays, CD8+ TIL are fully competent cytotoxic effector cells when subjected to strong activation signals via the TcR complex. In addition, these results imply that adoptively transferred CD4+ populations of TIL have in vivo biologic functions quite distinct from those of CD8+ populations and, further, that disparate clinical outcomes could reasonably be expected from the adoptive transfer of either population alone.

Antigens, Differentiation, T-Lymphocyte↗

PubMed1989

Role of thromboxane in interleukin 2-induced lung injury in sheep.

Interleukin (IL)-2 administration leads to respiratory dysfunction due to increased vascular permeability. This study examines the role of thromboxane (Tx)A2 in IL-2 induced lung injury in sheep with chronic lung lymph fistulae. This preparation enables evaluation of permeability prior to the development of gross edema. IL-2, 10(5) units/kg (n = 6), or its excipient control (n = 5) was given as an i.v. bolus over 2 min. After 2 h of IL-2 administration, plasma TxB2 increased from 168 to 388 pg/ml (P less than 0.05) and lung lymph TxB2 from 235 to 694 pg/ml (P less than 0.05). Mean pulmonary artery pressure (MPAP) rose from 13 to 29 mm of Hg (P less than 0.05) at 30 min and remained elevated for 4 h while the pulmonary artery wedge pressure was unchanged at 4 mm of Hg. Arterial oxygen tension (PaO2) fell from 88 to 77 mm of Hg (P less than 0.05). Lung lymph flow (QL) rose from 2.2 to 3.8 ml/30 min (P less than 0.05) at 1 h and to 6.4 ml/30 min at 3 h. This rise coincided with an increase in the lymph/plasma (L/P) protein ratio from 0.67 to 0.77 (P less than 0.05). In contrast, the non-IL-2-infused sheep (n = 3) recruitment of the lung vasculature by left atrial balloon inflation led to a rise in QL from 2.4 to 8.2 ml/30 min, whereas the L/P ratio declined from 0.62 to 0.25, suggesting that the protein-rich lymph flow after IL-2 administration reflected increased microvascular permeability. In further proof of an increase in permeability, IL-2 administration into sheep (n = 2) with an inflated left atrial balloon led, after a pressure-independent L/P protein ratio had been achieved, to an increase in L/P protein ratio and decrease in protein reflection coefficient. At 2 h after IL-2, the blood leukocyte count fell from 8156 to 4375/mm3 (P less than 0.05) primarily due to a 73% drop in lymphocytes. The platelet count declined from 292 to 184 x 10(3)/mm3 (P less than 0.05). Body temperature rose from 38.9-40.3 degrees C (P less than 0.05), and shaking chills were common. Pretreatment with the Tx synthetase inhibitor OKY 046 (n = 7) lowered baseline plasma and lymph TxB2 levels to 22 and 52 pg/ml (P less than 0.05) and prevented the IL-2-induced increase in plasma and lung lymph TxB2 (P less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1989

Activation of eosinophils in cancer patients treated with IL-2 and IL-2-generated lymphokine-activated killer cells.

Of 16 patients, a total of 13 who received IL-2 and autologous IL-2-generated lymphokine-activated killer LAK cells developed eosinophilia late during the course of treatment. To understand the direct or indirect effects of IL-2 on eosinophils, the physical and functional characteristics of the late-treatment eosinophils were compared to those of early-treatment and control eosinophils. Late-treatment eosinophils differed from early-treatment and control eosinophils in the following respects: they had somewhat reduced density, hypersegmented nuclei, eosinophil cationic protein converted from the storage form to the secretory form, and a greater than 200% increased ability to kill larvae of Schistosoma mansoni by an antibody-dependent mechanism (cytotoxic function). In vitro, IL-2 (1000 U/ml in medium as used to culture LAK cells) did not affect the cytotoxic function of eosinophils from cancer patients or from control subjects. However, LAK cell-conditioned medium enhanced the cytotoxic function of eosinophils from early-treatment cancer patients and from normal subjects by greater than 150%. Thus, eosinophils late in the course of IL-2/LAK cell treatment undergo physical changes and become functionally activated. The involvement of IL-2 in these changes is probably indirect, as an inducer of factors that enhance eosinophil function.

Antibody-Dependent Cell Cytotoxicity↗

PubMed1989

Ibuprofen causes reduced toxic effects of interleukin 2 administration in patients with metastatic cancer.

Metastatic cancer was treated with interleukin 2 and lymphokine-activated killer cells with the addition of the cyclooxygenase inhibitor ibuprofen in an attempt to reduce side effects in 13 patients (eight male and five female). Twenty-six patients treated with only interleukin 2 and lymphokine-activated killer cells formed the control group. After interleukin 2 administration, a significantly increased number of lymphokine-activated killer cells were transfused in ibuprofen-treated patients. Cytotoxic effects were not significantly different in the treated and untreated groups. With regard to cell phenotype, both groups of patients manifested significant activation of the immune system as measured by T10 and OK1a. Symptom scores were dramatically reduced in patients treated with ibuprofen. Temperature above 37 degrees C were rare. Ibuprofen did not significantly alter rate of response in this immunotherapy trial (38% vs 42%). Ibuprofen is now routinely used in all of our current immunotherapy trials.

Cells, Cultured↗

PubMed1989

Immunomodulatory effects of systemic low-dose recombinant interleukin-2 and lymphokine-activated killer cells in humans.

The adoptive immunotherapy of human cancer using lymphokine-activated killer (LAK) cells in combination with high-dose systemic recombinant interleukin-2 (rIL-2) has been associated with global changes in several hematological and immunological parameters while imposing profound toxicity on patients. We have evaluated an alternative LAK cell therapy utilizing low-dose systemic rIL-2 is also characterized by significant changes in immunological and hematological parameters, which are qualitatively similar to those induced by high-dose rIL-2. Low-dose systemic rIL-2, given by i.v. bolus, is cleared to baseline levels within 240 min of administration. The induction of lymphocytosis and eosinophilia, which has characterized other protocols, is also a feature of this protocol. In addition, low-dose systemic rIL-2/LAK cell immunotherapy results in increased peripheral blood mononuclear cell (PBMC) expression of T-cell activation markers such as OKIa, OKT10 and IL-2 receptor. PBMC sampled approximately 100 h after the final infusion of LAK cells demonstrated a statistically significant increase in their ability to kill natural killer (NK)-sensitive and NK-resistant cell lines such as K562 and Daudi compared to baseline values (P less than .05). These data suggest that rIL-2-based immunotherapy using low-dose rIL-2 is capable of inducing quantitative hematological and immunological changes while (in combination with LAK cells) retaining the ability to mediate tumor regression in vivo.

Adjuvants, Immunologic↗

PubMed1989

The rapid induction by interleukin-2 of pulmonary microvascular permeability.

The clinical use of interleukin-2 (IL-2) is limited by severe cardiopulmonary dysfunction. This study examines the mechanism of respiratory failure related to IL-2, using sheep with chronic lung lymph fistulae. Awake animals were infused with an intravenous (I.V.) bolus of IL-2 10(5) U/kg (n = 5) or its excipient (EXC) control (n = 3), every 8 hours for 4 to 5 days. Cardiopulmonary function was monitored daily for at least one 8-hour period. Within 2 hours after each IL-2 administration, mean pulmonary arterial pressure (MPAP) rose. On Day 1, the mean rise was from 13 to 26 mmHg (p less than 0.05), and on Day 5, to 29 mmHg (p less than 0.05). MPAP returned to baseline levels after 2-3 hours. Pulmonary arterial wedge pressure was unchanged from 4 mmHg. There were transient falls in arterial oxygen tension, from 88 to 77 mmHg on Day 1 and to 73 mmHg (p less than 0.05) on Day 5. Lung lymph flow (QL) rose from 2.4 to 6.8 ml/30 minutes (p less than 0.05) on Day 1, and from 4.7 to 10.2 ml/30 minutes (p less than 0.05) on Day 5, whereas the lymph/plasma protein ratio increased on Day 1 from 0.69 to 0.83 (p less than 0.05) and from 0.63 to 0.71 (p less than 0.05) on Day 5. This documents an increase in pulmonary microvascular permeability. Thromboxane (Tx)B2 levels increased transiently after each IL-2 injection in plasma from 195 to 340 pg/ml (p less than 0.05) and in lung lymph from 222 to 772 pg/ml (p less than 0.05) on Day 1, and to similar levels on Day 5. There was a progressive rise in cardiac output from 5.7 to 8.6 1/minute (p less than 0.05) during the 5 days of infusion. Systemic blood pressure did not change. Temperature rose from 39.1 to 41.2 C (p less than 0.05), and shaking chills were common. There was a progressive fall in leukocyte count, from 8.4 to 3.2 X 10(3)/mm3 (p less than 0.05) by Day 5, reflecting a 77% fall in lymphocytes. Lung lymph lymphocyte counts rose, and lymphocyte clearance increased.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1988

Adoptive immunotherapy of human cancer using low-dose recombinant interleukin 2 and lymphokine-activated killer cells.

The adoptive transfer of recombinant-methionyl human interleukin 2 (rIL-2)-activated autologous peripheral blood mononuclear lymphokine-activated killer (LAK) cells to cancer patients is being evaluated as an alternative to conventional cancer therapy. We have independently developed an alternative regimen to previously reported adoptive immunotherapy protocols using rIL-2 and LAK cells which features the prolonged administration of low-dose rIL-2 (30,000 units/kg) and an automated, entirely enclosed system of peripheral blood cell procurement, culture, harvest, and reinfusion of activated cells. The cell culture system was tested with a murine tumor model in which LAK cells generated in plastic culture bags were reinfused into tumor-bearing mice. Tumor regression was as effective with cells activated in the bags as in conventional culture flasks. Twenty-eight cancer patients were treated for 5 consecutive days with low-dose rIL-2, followed by leukapheresis, infusion of LAK cells, and prolonged IL-2 administration. At least 50% tumor regression was observed in 46% of all patients treated. These data imply that human peripheral blood mononuclear cells retain fully their capacity for rIL-2-induced activation and effector cell function under this alternative approach, and further, that a low-dose rIL-2 regimen with markedly reduced toxicities can be as effective as high-dose rIL-2 regimens if low-dose rIL-2 is given for a prolonged period of time following LAK cell infusion.

Adult↗

PubMed1988

Interleukin-2 initiates metabolic responses associated with critical illness in humans.

The cytokine interleukin-2 is a primary modulator of the immune response that occurs after infection, trauma, and transplant rejection, yet its role as a mediator of associated metabolic changes in surgical illness is unknown. We studied clinical and metabolic responses in eleven tumor-bearing humans with normal renal and hepatic function receiving bolus intravenous (I.V.) interleukin-2 (30,000 U/kg). Additional subjects (n = 6) were pretreated with the cyclooxygenase inhibitor, ibuprofen (1600 mg, orally), before interleukin-2 administration. Serial measurements were made of vital signs, symptoms, hematology, and plasma concentrations of pituitary and stress hormones and selected cytokines. Administration of interleukin-2 resulted in fever, tachyacardia, "flu-like" symptoms, and neurohormonal elaboration. The responses observed were quantitatively similar to those that occurred after endotoxin administration in healthy subjects (n = 13), but differed in the following manner: 1) the onset of fever and endocrine changes occurred after a longer latent interval (180-240 minutes vs. 60-90 minutes after endotoxin), 2) peak responses after the administration of interleukin-2 also occurred later, 3) no increased circulating tumor necrosis factor was detected after administration of interleukin-2 (peak plasma concentration was greater than 35 pg/ml vs. 270 +/- 70 pg/ml after endotoxin administration), and 4) administration of interleukin-2 but not of endotoxin was associated with increased circulating concentrations of gamma interferon (peak plasma concentration 1.7 +/- 0.2 NIH U/ml vs. less than 0.1 NIH U/ml after endotoxin administration). Fever and neurohormonal responses after interleukin-2 administration were greatly attenuated by ibuprofen administration. Interleukin-2 induces other cytokines that exert their effects largely through the cyclooxygenase pathway. Interleukin-2 may be an important signal, initiating the integrated host responses to infection and injury.

Adrenocorticotropic Hormone↗

PubMed1988

Efficacy of specimen radiography of clinically occult noncalcified breast lesions.

Breast biopsy specimen radiography is required to ensure the accurate removal of clinically occult lesions discovered by mammography. Although used routinely for calcified lesions, it has not been widely accepted for those abnormalities that do not contain calcium. To determine the efficacy of film-screen specimen radiography for confirming the presence of clinically occult, noncalcified lesions, we undertook a prospective study of 104 specimen radiographs obtained after mammographically guided hookwire localization and planned excision of these lesions. Ninety-seven (93%) of the excised abnormalities were visualized on specimen mammograms. Malignancy was found in 22 (21%) of them. Thirty-five percent of the specimen radiographs showed better anatomic detail of the lesion, 48% showed the same detail, and 16% showed less detail than the original mammograms. Specimen radiographs failed to show the lesion in only seven cases. Five of the seven were true-negative specimen radiographs, making the efficacy rate 98%. Film-screen specimen radiography of clinically occult, noncalcified lesions is a highly effective procedure for correctly identifying the presence of a mammographic abnormality.

Breast Diseases↗

PubMed1988

A new regimen of interleukin 2 and lymphokine-activated killer cells. Efficacy without significant toxicity.

Adoptive immunotherapy with high-dose interleukin 2 and lymphokine-activated killer (LAK) cells has proved to be successful in the treatment of some patients with metastatic cancer, but not without a significant degree of associated toxic effects. The primary goal of this study was to substantially reduce the toxicity of this complex and expensive treatment, while maintaining or improving efficacy. To this end, 29 patients were treated with LAK cells in conjunction with a low-dose regimen of interleukin 2 and a prolonged period of administration following LAK cell infusion. This protocol resulted in a considerable reduction in toxicity, as compared with that described in previous studies, without compromising the efficacy. This study offers further confirmation that adoptive immunotherapy of metastatic cancer can be clinically beneficial to patients for whom no other effective therapy is presently available.

Adult↗

PubMed1984

Phase I study of the adoptive immunotherapy of human cancer with lectin activated autologous mononuclear cells.

In previous in vitro studies, the authors showed that phytohemagglutinin (PHA) stimulated peripheral blood lymphocytes (PBL) from cancer patients to generate cells that were lytic for fresh autologous tumor but not for lymphocytes or lymphoblasts. Thus, after IRB approval, a phase I clinical protocol was instituted in cancer patients who had failed all other therapy to determine the toxicity and effects, in vivo, of the infusion of large numbers of such PHA activated autologous PBL. Ten patients were treated on the protocol, six with sarcoma, one with melanoma, and three with colorectal cancer. Up to a total of 1.7 X 10(11) PBL were obtained from 7 to 15 successive leukaphereses, the cells from each leukapheresis being incubated in vitro in medium containing PHA and human AB serum for 2 days and then reinfused following the next leukapheresis 2 days later. Toxicity encountered included fever and chills in 10/10 patients, headaches in 5/10, nausea and vomiting in 3/10, and requirement for erythrocyte transfusion in 8/10. No evidence for autoimmune disease, abnormal serum chemical or coagulation studies, or pulmonary emboli was found. 111Indium trafficing studies showed distribution of infused cells mainly to the spleen and liver, with some accumulation in the lungs and tumor especially after repeated infusions. In 9/10 patients, activated PBL were detected in the peripheral circulation by the sixth leukapheresis. Evidence for this was found by assaying the incorporation of tritiated thymidine (3H-Tdr) into, and lysis of fresh tumor cells by, unstimulated PBL from successive leukaphereses. No tumor regression was seen in these patients with bulk disease. These studies demonstrated that large numbers of PHA-activated PBL can be safely obtained and infused into humans, achieving an increase in the number of circulating activated cells with evidence of migration of cells to tumor, lungs, liver and spleen. Further studies of the use of activated lymphocyte infusion in conjunction with chemotherapy in humans are in progress.

Adult↗

PubMed1984

Adoptive immunotherapy of established syngeneic solid tumors: role of T lymphoid subpopulations.

We have investigated the subpopulations of T cells necessary to mediate the cure of established tumors in two models of successful adoptive immunotherapy. In C57BL/6 mice bearing palpable and disseminated FBL-3 lymphoma, both Lyt-1+ and Lyt-2+ cells played a major role in mediating the regression and permanent cure of mice, whereas in BALB/c mice bearing the Meth A sarcoma the adoptive transfer of Lyt-1+2+ cells played a major role in mediating the regression of tumors and the curing of disease. Identical experiments performed in hybrid (BALB/c X C57BL/6) mice yielded similar results, further supporting our initial observation and indicating that in these two adoptive transfer model systems it is the tumor and not the variable expression of Lyt antigens by the host that determines which T cell subpopulation is required to cure mice of tumors.

Animals↗

PubMed1983

Clonal analysis of the lymphoid cells mediating skin allograft rejection. Mediation of graft rejection in vivo by cloned Lyt-1+2- proliferative, noncytotoxic long-term cell lines.

Studies were undertaken to elucidate the cellular basis of skin allograft rejection mediated by long-term cultured cell lines and clones. The adoptive transfer, in vivo, of in-vitro-sensitized cells, from B6AF1 anti B10.BR or from C57BL/6 anti DBA/2 cultures, and expanded eight-fold to ten-fold for one week in lectin-free interleukin 2 (LF-IL-2) were able to mediate specific skin allograft rejection. These same cells lost the ability to mediate accelerated skin graft rejection when they were expanded more than 100-fold during three weeks of culture in LF-IL-2 even though these cultures mediated high levels of specific in vitro cytotoxicity for the appropriate allosensitizing cells. When Lyt-2+ cells were depleted using monoclonal antibodies and complement prior to in vitro sensitization and expansion in LF-IL-2, these cells lines retained the ability to mediate skin allograft rejection in vivo when expanded more than 100-fold for three culture generations in vitro. These latter lines were greatly enriched for Lyt-1+2- cells and had little or no cytolytic activity, but they retained specific in vitro proliferative responses to the sensitizing alloantigen. Several Lyt-1-2+ cloned long-term lymphoid cell lines with high levels of specific cytolytic activity against the sensitizing alloantigen were derived and none was capable of mediating the accelerated rejection of skin grafts in vivo. However, cloned lymphoid cell lines that were phenotypically Lyt-1+2- and were capable of proliferating when in contact with specific alloantigen, but were not cytolytic, were capable of mediating the accelerated rejection of skin grafts in vivo both in irradiated mice and in nude mice. These studies demonstrate that skin allograft rejection can be mediated by Lyt-1+2- cell lines with specific in vitro proliferative activity to alloantigen although Lyt-1-2+ cell lines with cytolytic but not proliferative activity to alloantigen in vitro are ineffective in mediating graft rejection in vivo. Specific proliferative activity and no cytolysis appears to be a good in vitro correlate of the in vivo activity of long-term cultured cell lines.

Animals↗