Reactivity Of Peripheral Blood Lymphocytes Research Articles

Vaccinia virus as a vaccine delivery system for marsupial wildlife

Vaccines based on recombinant poxviruses have proved successful in controlling diseases such as rabies and plague in wild eutherian mammals. They have also been trialled experimentally as delivery agents for fertility-control vaccines in rodents and foxes. In some countries, marsupial mammals represent a wildlife disease reservoir or a threat to conservation values but, as yet there has been no bespoke study of efficacy or immunogenicity of a poxvirus-based vaccine delivery system in a marsupial. Here, we report a study of the potential for vaccination using vaccinia virus in the Australian brushtail possum Trichosurus vulpecula, an introduced pest species in New Zealand. Parent-strain vaccinia virus (Lister) infected 8/8 possums following delivery of virus to the oral cavity and outer nares surfaces (oronasal immunisation), and persisted in the mucosal epithelium around the palatine tonsils for up to 2 weeks post-exposure. A recombinant vaccinia virus construct (VV399, which expresses the Eg95 antigen of the hydatid disease parasite Echinococcus granulosus) was shown to infect 10/15 possums after a single-dose oronasal delivery and to also persist. Both parent vaccinia virus and the VV399 construct virus induced peripheral blood lymphocyte reactivity against viral antigens in possums, first apparent at 4 weeks post-exposure and still detectable at 4 months post-exposure. Serum antibody reactivity to Eg95 was recorded in 7/8 possums which received a single dose of the VV399 construct and 7/7 animals which received triple-dose delivery, with titre end-points in the latter case exceeding 1/4000 dilution. This study demonstrates that vaccinia virus will readily infect possums via a delivery means used to deploy wildlife vaccines, and in doing is capable of generating immune reactivity against viral and heterologous antigens. This highlights the future potential of recombinant vaccinia virus as a vaccine delivery system in marsupial wildlife.

Direct evidence that naturally occurring mutations within hepatitis B core epitope alter CD4+ T‐cell reactivity

Exacerbations of chronic hepatitis B have been associated with accumulation of mutations in the HBV core gene, with amino acid (aa) substitutions clustering between aa 50 and 69. This region of the nucleocapsid protein is known as an immunodominant epitope for CD4+ T-lymphocytes, however the impact of these mutations on T-cell reactivity has not been investigated. For this purpose, we undertook fine mapping of the reactivity of peripheral blood lymphocytes, isolated from patients with acute (n = 8) or chronic hepatitis B (n = 10), against a panel of branched synthetic peptides. The peptide aa sequences corresponded to the wild type HBV (aa 50-69), or contained 1-3 aa changes derived on the basis of naturally occurring mutations. In four of eight patients with acute hepatitis B the wild type peptide 50-69, which corresponded to the core gene sequence of HBV present in these patients, induced a strong T-cell proliferative response. In the same cases, the T-cell response to the mutant peptides was altered at various degrees, depending on the number and the position of aa changes. The most pronounced inhibition of CD4+ T-cell response (between 44 and 92%) was caused by a peptide ligand with two aa substitutions at positions 64 and 67. These results demonstrate that mutations within immunodominant epitopes of the HBV nucleocapsid can affect the CD4+ T-lymphocyte reactivity, which may have a role for the accumulation of certain HBV strains after hepatitis flares during the course of chronic HBV infection.

Enzyme-linked immunosorbent spot assay analysis of peripheral blood lymphocyte reactivity to donor HLA-DR peptides: potential novel assay for prediction of outcomes for renal transplant recipients.

Chronic allograft dysfunction, which is the most common cause of late allograft failure, is in part caused by an ongoing immune response orchestrated by T lymphocytes primed by the indirect pathway of allorecognition. The low frequencies of such T cells have made it difficult to study indirect alloreactivity by using currently available assays. The development of a sensitive, clinically useful method of measuring indirect alloreactivity among human renal transplant recipients was thus attempted. Furthermore, in a pilot immunologic study, the contribution of the indirect pathway was studied in two groups of renal transplant recipients, i.e., patients with no prior acute rejection episodes and stable renal function ("stable" patients) and patients with at least one previous episode of biopsy-proven acute rejection, who were thus at risk for the development of chronic rejection ("high-risk" patients). The frequencies of type 1 T helper (interferon-gamma-producing) and type 2 T helper (interleukin-5- and -10-producing) peripheral blood lymphocytes reactive with a panel of synthetic peptides (corresponding to sequences from donor HLA-DR molecules) were determined for renal transplant recipients and normal control subjects by using an enzyme-linked immunosorbent spot assay (ELISPOT). Among recipients of DR-mismatched allografts, a cut-off value of 60 interferon-gamma spots/10(6) cells significantly (P = 0.02) separated stable patients (creatinine concentration, 1.1 +/- 0.3 mg/dl) from high-risk patients (creatinine concentration, 2.3 +/- 1.7 mg/dl). This is the first demonstration that the enzyme-linked immunosorbent spot assay can be used to monitor indirect alloreactivity to donor HLA-DR peptides among renal transplant recipients. These data provide the rationale for the prospective study of indirect alloreactivity among transplant recipients, to allow predictions of which patients would be at risk for the development of chronic rejection and thus allow appropriate planning of future interventions.

Oral Infection of Ferrets with Virulent Mycobacterium bovis or Mycobacterium avium: Susceptibility, Pathogenesis and Immune Response

Ferrets are important wildlife reservoirs of tuberculosis in New Zealand, where they acquire infection primarily through scavenging infected carrion. In the present study, groups of laboratory-reared ferrets were infected orally with 5×106colony-forming units of Mycobacterium bovis or Mycobacterium avium. Body weight and tuberculin-specific immune reactivity were monitored at intervals (pre-infection, and 4 and 20 weeks post-infection) and animals were killed at 20 weeks post-infection for post-mortem, histopathological and bacteriological examinations. Weight loss was significantly greater in M. bovis -infected than in M. avium -infected ferrets. M. bovis, unlike M. avium, sometimes produced gross necrotic lesions in the mesenteric lymph nodes. M. bovis invariably produced microscopical foci of mycobacterial infection or tissue necrosis typical of tuberculosis, whereas M. avium did so in only one of nine animals. Mycobacteria were recovered from the lymphatic tissues of all M. bovis -infected ferrets but from only five of nine M. avium -infected animals; and the mean bacterial burdens of the lymph nodes of the head and intestinal regions were >10-fold and >100-fold greater, respectively, for M. bovis -infected than for M. avium -infected animals. M. bovis, unlike M. avium, evoked tuberculin-specific peripheral blood lymphocyte reactivity and serum antibody responses.

Reactivity of peripheral blood lymphocytes to oxidized low-density lipoprotein: a novel system to estimate atherosclerosis employing the Cellscan.

The assumption that atherosclerosis involves an autoimmune response to oxidized LDL (oxLDL) is based on the presence of immunocompetent cells and immunoglobulin deposition in the atherosclerotic lesions by successful immunomodulation of the atherosclerotic process and by inhibition of experimental atherosclerosis by antioxidants. The Cellscan system is a multiparameter laser-based static cytometer that enables repeated monitoring of the fluorescence intensity (FI) and polarization (FP) of individual living cells. Analysis of intracellular fluorescein fluorescence polarization (IFFP) has previously been used to define activated lymphocyte population. In this study, the Cellscan apparatus has been used to monitor cellular response to oxLDL in patients with atherosclerosis and in controls. The FI and FP of fluorescein diacetate (FDA)-labeled peripheral lymphocytes were measured following exposure to oxLDL in vitro. Using cluster analysis we were able to identify subpopulations of cells that were characterized by their FI and FP. Forty-two subjects were studied: 22 patients with severe coronary heart disease and 22 control individuals, either healthy or with other diseases. Fluorescence intensity of fluorescein-labeled peripheral blood lymphocytes (PBL) was markedly decreased upon exposure to high doses (> 25 micrograms/ml) of oxLDL concurrently with an increase in FP. A specific and dose-dependent reduction in FP of the high-intensity cell subpopulations, accompanied by higher FI, was evident in patients with ischemic heart disease upon exposure to low doses of oxLDL (up to 25 micrograms/ml). Maximal depolarization was shown upon triggering with 2 micrograms/ml oxLDL. The polarization ratio (the mean polarization value of the specific cell population with and without activation) obtained for patients' lymphocytes was significantly lower (p < 0.01) than that of the control group (0.936 +/- 0.05 and 1.028 +/- 0.055, respectively). These data suggest that PBL from patients with active ischemic heart disease show an increased reactivity to oxLDL. A 73% positivity rate was found for ischemic heart disease patients compared with 5% in the control subjects. One of the future prospects of this study might be the advent of a simple and rapid noninvasive test that could assess the extent of atherosclerosis, and possibly even the response to therapy, by monitoring the reactivity of PBL to oxLDL.

Autoimmune Deafness is not Related to Hyperreactivity to Type II Collagen

The pathogenic role of anti-type II collagen was analysed in a variety of hearing losses, in age-matched controls and in different autoimmune diseases. The immune reactivity of peripheral blood lymphocytes to type II collagen was studied by the degree of proliferation measured as the incorporation of bromodeoxyuridine in cultured lymphocytes. The anti-type II collagen antibodies showed a very low incidence in the hearing loss group. Lymphocytes of otosclerosis, Meniere's disease and other sensorineural deafness patients proliferated in response to concanavalin A and to type II collagen to a lower extent than peripheral blood lymphocytes from healthy controls. Nonetheless, these differences were not statistically significant. The immune hyperreactivity to type II collagen cannot explain the autoimmune mechanism of hearing losses. Humoral and cellular hyperreactivities to inner ear proteins different from type II collagen, could explain the autoimmune mechanism of deafness.

Genes of chicken MHC regulate the adherence activity of blood monocytes in Rous sarcomas progressing and regressing lines

The influence of the chicken major histocompatibility (B) complex (MHC) on the adherence potential of monocyte-derived macrophages was examined using the congenic chicken lines CB and CC. These lines represent well-defined genetic models for the study of resistance (CB) or susceptibility (CC) to the progressive growth of Rous sarcomas. Using a monoclonal antibody specific for chicken monocytes/macrophages, CB and CC chickens were shown by flow cytometry analyses to have similar proportions of peripheral blood monocytes. However, when the glass-adherence potential of these cells was compared during incubation in tissue culture medium over 24, 48 and 72 h at 40°C, significant differences were seen between cells from these two inbred lines. After 24 and 48 h, glass-adherence by CB cells was 2–3 fold higher than that of CC cells. After 72 h this difference decreased to 1.5 fold. At 24 and 48 h, the adherent CB macrophages also appeared about 1.5 times larger than those of CC chickens. Genetic analysis using F 1 hybrids (CB×CC) showed that this trait is regulated by a dominant gene that segregates with the B12 haplotype in the backcross generation F 1×CC. From the results obtained with the recombinant congenic lines CB.R1 and CC.R1, we conclude that the gene regulating adherence potential is localized within the B-F/L region of the chicken MHC. About 50% of adherent cells were able to phagocytose opsonised FITC-labelled Zymosan particles. The level of nitric oxide production in vitro by CB and CC macrophages was equal. The importance of cells of the mononuclear phagocyte system for the response to Rous sarcoma virus (RSV) infection was studied in CB chickens using the anti-macrophage agents silica, carrageenan, and Cl 2MDP, encapsulated in liposomes. In those chickens treated with silica and carrageenan, we observed progressive growth of RSV-induced tumors. The graft-versus-host reactivity of peripheral blood lymphocytes (PBL) of treated chickens was comparable to controls. In vitro nitric oxide production by macrophages from silica-treated chickens was higher than by macrophages from untreated controls.

Use of cryopreserved lymphocytes for assessment of the immunological effects of interferon therapy in renal cell carcinoma patients

Experiments were designed to assess whether cryopreserved PBL could be used to monitor the immunological effects of IFN-α therapy in renal cell carcinoma (RCC) patients. It was found that programmed freezing and thawing of peripheral blood lymphocytes (PBL) from normal blood donors did not substantially change lymphocyte subset proportions and that cryopreserved PBL were able to proliferate in response to IL-2. It was also possible to activate the cytolytic activity of frozen PBL, and the frozen leukocytes did not lose their ability to secrete IFN-γ after PHA activation. We have used these findings to investigate the immunological effects of IFN-α therapy in RCC patients. Cryopreservation of PBL samples collected from various patients over a period of 9–14 months enabled us to compare the in vitro reactivity of PBL from individual RCC patients repeatedly and under standard conditions. It was found that IL-2-induced proliferative responses of PBL from IFN-α non-responders, collected prior to IFN-α therapy, were significantly decreased as compared to those from normal blood donors. The proliferative responses of PBL from IFN-α responders, collected prior to IFN-α therapy, did not substantially differ from normal controls. Culture of PBL from IFN-α responders for 3 days in IFN-α-containing medium increased their lytic activity towards RCC targets, whereas no such increase was observed with non-RCC targets or using PBL from IFN-α non-responders or PBL from normal blood donors. Enzyme-linked immunospot (ELISPOT) assays performed with cryopreserved lymphocytes from IFN-α non-responding RCC patients, collected prior to IFN-α therapy, revealed a substantially decreased ability to secrete IFN-γ, as compared to IFN-γ secretion of PBL from IFN-α responders or normal blood donors.

IFN-ALPHA THERAPY OF RENAL-CELL CARCINOMA - DEFECT OF LYMPHOCYTE SENSITIVITY TO MITOGENIC AND ACTIVATING CYTOKINE SIGNALS IN PATIENTS NOT-RESPONDING TO THERAPY

The present prospective study was designed to assess whether the renal cell carcinoma (RCC) patients treated with recombinant interferon alpha (IFN alpha), whose tumours respond (responders) and do not respond (non-responders) to IFN alpha therapy, differ with regard to in vitro sensitivity of peripheral blood lymphocytes (PBL) to interleukin 2 (IL-2), IFN alpha, and IFN gamma signals prior to therapy. Twenty-one patients with advanced RCC after nephrectomy, 15 responders and 6 non-responders, were entered into a protocol. The protocol involved isolation and freezing of PBL samples followed by IFN alpha treatment of patients, assessment of proliferative and activating PBL responses, and evaluation of the therapeutic results. Freezing of PBL samples allowed us to compare the in vitro reactivity of PBL from individual RCC patients, repeatedly and under standard conditions. Substantial differences in proliferative responses to the mitogenic IL-2 signal of PBL derived from IFN alpha responders and nonresponders were found. Whereas the IL-2-induced proliferative responses of PBL from normal blood donors and IFN alpha responders were comparable, the proliferative responses of PBL from IFN alpha non-responders were significantly decreased, suggesting an immune dysfunction in non-responders. Cultivation of PBL from RCC patients in medium supplemented with IFN alpha increased the lytic activity of PBL from IFN alpha responders directed against RCC targets; no such increase could be observed with non-RCC targets, with PBL from IFN alpha non-responders, or with PBL from normal blood donors. Detection of phytohaemagglutinin (PHA)-stimulated IFN gamma secretion by PBL at the single cell level using enzyme-linked immunospot (ELISpot) assay revealed that the ability to produce IFN gamma was substantially decreased in IFN gamma non-reponders, as compared to IFN alpha responders and to normal blood donors.

Prophylactic therapy with OKT3 does not affect donor specific reactivity of peripheral blood lymphocytes from heart transplant recipients

To avoid the nephrotoxic effect of high-dose cyclosporin A (CsA) immediately post-transplant, heart transplant recipients received as prophylactic therapy intravenous OKT3 for seven days instead of intravenous CsA. Patients receiving OKT3 were compared with patients receiving CsA with respect to specific proliferation and cytotoxicity of their peripheral blood mononuclear cells (PMNC) against donor antigens, at different times within three months post-transplant. No effect of the initial immunosuppressive therapy was observed on these parameters. Acute rejection did not induce a consistent effect on the relative proliferation. PMNC from patients who experienced one or more rejection episodes showed a decrease in donor specific relative proliferative response in time after transplantation, while nonrejectors did not or only slightly, independent of the initial immunosuppressive protocol. Within the group of patients not receiving OKT3, this effect of rejection reached significance. In conclusion, no effect of prophylactic OKT3 therapy compared with prophylactic CsA therapy was observed on donor reactivity of PMNC in vitro during the subsequent three to four months post-transplant.

Opposite effects of mild and severe stress on in vitro activation of rat peripheral blood lymphocytes

The effects of short-term handling and different durations of immobilization on serum levels of catecholamines, ACTH, prolactin, and corticosterone and in vitro functions of lymphocytes were examined in rats. The results show that changes in the immune response of peripheral blood lymphocytes (PBL) depend on the intensity of the stressor: Short (1 min) handling of cannulated rats induced an enhanced stimulation of PBL to respond to T and B cell mitogens, whereas immobilization of the same animals led to suppression, dependent on the time this stressor was applied. The decrease in the mitogen reactivity of PBL after 120 min of immobilization was reversible within 24 h, and could be largely prevented by adrenalectomy, confirming that factors released by this gland are mainly responsible for immunosuppression. In contrast to PBL, spleen cells showed an enhanced mitogen response to immobilization and adrenalectomy, indicating that the immune response is differently regulated in the various compartments of the immune system. Possible correlations of the various effects with changes in stress hormone levels are discussed.

Influence of immunotherapy on the cellular immunity of unexplained recurrent aborters

Changes in lymphocyte subsets in whole blood were analyzed sequentially by flow cytometry with an automated leukocyte differential system in 15 patients with unexplained recurrent spontaneous abortions, each of whom underwent vaccination(s) with her husband's lymphocytes. Mitogen responses of peripheral blood lymphocytes (PBL) were also examined in these patients. The reactivity of PBL against mitogens revealed no significant change in each patient before and after vaccination(s) with her husband's lymphocytes. The CD4:8 ratio was observed to decrease significantly during 22 and 28 days after the first vaccination with a significant increase in the percentage of T suppressor-cytotoxic (CD8) cells. This change was also observed after the second vaccination. The percentages of other subsets did not change significantly after vaccination(s). In 11 patients out of 15, the pregnancy continued successfully and correlated with a predominance of Ts/c (CD8) over TH/1 (CD4) cells in the first trimester. These changes in lymphocyte subsets may indicate the induction of immune enhancing mechanisms and it is suggested that continuation of the predominance of Ts/c cells induced by immunotherapy might be important for the successful maintenance of pregnancy.

In vitro modulating effects of porcine immunoglobulin G on mitogens-induced lymphocyte response in precolostral, suckling and weaned piglets

The influence of allogeneic IgG on in vitro reactivity of peripheral blood lymphocytes (PBL) of neonatal colostrum-deprived piglets as well as of suckling and weaned piglets was studied. PBL were preincubated with purified allogeneic IgG for 24 h before their ability to respond to PHA, Con A or PWM was tested. PBL of precolostral piglets pretreated with allogeneic IgG exhibited higher response to PHA ( P < 0.01) than untreated control cells. An increased response of PBL treated with IgG was also observed in suckling piglets as compared to their respective control cells ( P < 0.01). Responsiveness of PBL treated with IgG to PWM was suppressed. No differences in response to Con A regardless of the sources of lymphocytes was observed as compared to IgG untreated controls. The results suggest that pretreatment of lymphocytes of piglets with allogeneic IgG modulates their reactivity to mitogens, suppressing the response to PWM and stimulating the response to PHA, respectively.

Qualitative analyses of cellular immune functions in equine infectious anemia show homology with AIDS

Equine infectious anemia (EIA) is a disease caused by a lymphocytotropic lentivirus which belongs to the same subfamily as HIV. Because of the very close relationship of their predicted gag and pol gene products and similarities in clinical manifestations of the disease, EIA served as a model to study immunological events involved in the host defence against lymphocytotropic viral infections. The existence of antibody dependent cell-mediated cytotoxicity against autologous EIA virus infected lymphoblasts was demonstrated in vitro at the beginning of an acute attack of the disease. Cytotoxic activity was not found or was very low during chronic infection. Reactivity of peripheral blood lymphocytes to Protein A and allogeneic cells in vitro was significantly suppressed in the presence of an acute serum, while the reactivity to PHA was at the normal level. These results suggest that cellular immune mechanism(s) are also involved in removal of EIA virus infected cells as has been reported recently in HIV-1 infected individuals and that EIAV and HIV immunopathogenesis show similarities in affecting the immune response of the host.

Influence of the Hal locus and standardized stress on antibody response and in vitro reactivity of peripheral blood lymphocytes in pigs

The antibody response to two Escherichia coli antigens (0149 and K88) and the in vitro reactivity of peripheral blood lymphocytes (PBLs) to pokeweed mitogen was studied in growing pigs after exposure to varying degrees of stress. Three experimental groups were used; transportation by lorry, transportation by lorry after a previous injection of a tranquillizing drug (amperozide), no transportation but an amperozide injection. Another group was used as a control group. This group was not transported and received no amperozide injection. The animals were the offspring of boars and sows heterozygous ( Nn) for the Hal gene, and all 3 Hal genotypes ( NN, Nn, nn) were identified and could thus be compared within litters. Immediately after the experimental treatment, the highest PBL reactivity was found for the amperozide-treated animals and for the non-transported animals, with no differences in reactivity between Hal genotypes. Two weeks later, the treatments caused different effects on pigs of the 3 Hal genotypes, both with regard to the PBL reactivity and the IgG response to 0149. The NN pigs had a higher PBL reactivity than the nn pigs for all treatments except the ‘drug + transport’ class where the reverse rank order was found. The NN pigs also had a higher IgG response to 0149 than the nn pigs in the ‘drug + no transport’ class. The amperozide treatment was followed by a higher PBL reactivity in non-transported NN pigs and in transported nn pigs. The amperozide-treated non-transported NN pigs also had a higher IgG response to 0149. The highest PBL reactivity and IgG response to 0149 were found mainly in pigs with the lowest cortisol levels. Pronounced differences between litters were found for both the antibody response and the PBL reactivity.

Transmission of simian acquired immunodeficiency syndrome with a type D retrovirus: Immunological aspects

Simian acquired immunodeficiency syndrome (SAIDS) was transmitted to four of four rhesus macaques with blood from rhesus macaques naturally infected with a type D retrovirus, simian retrovirus-2 (SRV-2). Three of the four blood recipients died with SAIDS at 13, 15, and 26 weeks postinoculation. The fourth animal is alive with SAIDS. All four test monkeys became viremic and produced antiviral antibody. None of the inoculated monkeys produced measurable neutralizing antibody to SRV-2. The survivor produced higher levels of antiviral antibody than the monkeys that died. Phytohemagglutinin and concanavalin A reactivity of peripheral blood lymphocytes was depressed from weeks 6 to 12 after innoculation. Clinical findings included development of splenomegaly in all four monkeys, and diarrhea in two monkeys. Blood counts remained within the normal range except for a depression in the number of polymorphonuclear lymphocytes in two monkeys. Hematocrits were decreased in two monkeys just prior to their death. All four test monkeys developed lymph node atrophy and bone marrow hypoplasia. Total proteins and immunoglobulin production were normal. This report provides evidence that SRV-2, as well as other type D retroviruses, causes SAIDS in macaque species.

消化器癌における末梢血および癌所属リンパ節リンパ球の癌選択的幼若化能について

消化器癌における末梢血および癌所属リンパ節リンパ球の癌選択的幼若化能について

Pulmonary and immune responses to a Thermoactinomyces vulgaris antigen respiratory sensitization in C57BL/6J mice

Lung histology and immune responses were followed during a six month period of Thermoactinomyces vulgaris antigen respiratory sensitization of C57BL/6J mice. Histologically, the mice developed a picture resembling the subacute stage of human farmer's lung. At the early phase of the sensitization a T-cell response was seen in vitro, characterized by an increased spleen but no peripheral blood lymphocyte reactivity to T-cell mitogens at the same time as increased reactivity to the sensitizing antigen was detected. An increase in the percentage of T-cells in the lavage cell population was also seen. The sensitization was associated with the appearance of both IgG and IgA antibodies in lavage and in serum of which lavage IgA antibodies were found to follow the course of the sensitization most accurately. The increase in antibody titers was not, however, reflected in lavage cytology or in lymphocyte reactivity. The results suggest that different immunological reactions may be predominant at different stages of farmer's lung.

Study of Tumor-Associated Blastogenic Reactivity of Peripheral Blood Lymphocytes and Regional Lymph Node Lymphocytes in Human Malignancy

The in vitro blastogenic responses of peripheral blood lymphocytes (PBL) and the regional lymph node lymphocytes (LNL) from cancer patients to autochthonous tumor cells (TC) were investigated. A positive lymphocyte response to autochthonous TC was observed in 23% (6/26) for PBL and 34% (9/26) for LNL. There seemed to be an inverse relationship between PBL and LNL responses to autochthonous TC. There was no significant difference between PBL and LNL responses to autochthonous TC according to the primary site of the tumor. Analysis of PBL and LNL responses to autochthonous TC according to the extent of the disease revealed that LNL were definitely more reactive than PBL in patients with a primary tumor that had not spread to the regional lymph nodes.

Sequential in vitro reactivity of lymphocytes from patients with cervical squamous malignancy in a cytotoxicity assay

The in vitro reactivity of peripheral blood lymphocytes from patients with cervical squamous malignancy was prospectively followed over a relatively long period of time. In 12 of 14 patients with preinvasive cervical lesions, reactivity was present at the time of initial diagnosis. Three months after treatment, reactivity was still present in only one of 12 (8%). Six months after the treatment, no significant reactivity could be detected in any of them. In the group with invasive squamous cell carcinoma, four patients developed recurrence in the course of follow-up. Reactivity was present in all at the time of initial diagnosis. In three of them, cytotoxic reactivity was retained up to the time of death in spite of clinical deterioration. One lost the reactivity after the third course of chemotherapy. Fourteen patients with invasive squamous cell carcinoma were followed up to 24 months without any evidence of recurrence. In all of this group, the cell-mediated cytotoxicity was nonreactive at 9 months. We conclude that: (1) patients with squamous cell carcinoma of the cervix demonstrated cell-mediated immune responses which disappeared 3 to 9 months after effective treatment (apparent cure) and (2) with persistent disease, in spite of marked clinical deterioration and inanition, cell-mediated cytotoxicity was demonstrable until the time of death.