Amount Of Natural Killer Cells Research Articles

Ex vivo expansion of natural killer cells for hematological cancer immunotherapy: a systematic review and meta-analysis.

The present systematic review aimed to investigate natural killer (NK) cell ex vivo expansion protocols within the scope of clinical trials targeting hematological cancer and to conduct a meta-analysis to assess the effect of NK cell infusion on survival. Research articles of clinical studies in which cell products produced by ex vivo expansion, consisting of a certain amount of NK cells and infused to patients with hematological cancer, were included in the systematic review. We conducted a proportion analysis with random effects for product purity and viability values. Studies having control groups were included in the survival meta-analysis. Among 11.028 identified records, 21 were included in the systematic review. We observed statistically significant heterogeneity for viability (I2 = 97.83%, p < 0.001) and purity values (I2 = 99.95%, p < 0.001), which was attributed to the diversity among isolation and expansion protocols. In addition, the survival meta-analysis findings suggested that NK cell therapy favors disease-free survival (DFS) of patients with myeloid malignancies but limited to only two clinical studies (odds ratio = 3.40 (confidence interval:1.27-9.10), p = 0.01). While included protocols yielded cell products with acceptable viability, the utility of immunomagnetic methods; feeder cells such as K562 expressing membrane-bound IL15 and 4-1BBL or expressing membrane-bound IL21 and 4-1BBL might be preferable to achieve better purity. In conclusion, NK cell therapy has a potential to improve DFS of patients with myeloid malignancies.

Сравнительная характеристика эффективности применения гексапептида и азоксимера бромида в комплексном лечении детей с вторичным пиелонефритом

Objective. Secondary pyelonephritis occurs in children in a distant period after adequate surgical correction of congenital obstructive pathology due to long-lasting shifts of immunologic resistance. Patients and methods. The authors treated 80 children aged 5 to 15 years with secondary pyelonephritis with preserved renal function against hydronephrosis, megaureter, vesicoureteral reflux. Immunological parameters were assessed at the child's admission to the hospital and 3 months after in-patient treatment. Initial changes of immunological reactivity in the patients substantiated the treatment tactics. Group 1 consisted of 40 patients, whose complex treatment included the immunomodulator, the active ingredient hexapeptide; Group 2 consisted of 40 patients, whose complex treatment included the tablet preparation of Polyoxidonium, the active ingredient azoximer bromide. Results. Inclusion of and azoximer bromide in treatment of children with secondary pyelonephritis was found to lead to increasing of IgA, IgG, B-cells amount, phagocytosis rate and natural killer number; and to decreasing of the amount of T-lymphocytes and CD4-cells. Inclusion of hexapeptide in treatment of secondary pyelonephritis in children increases level of phagocytic activity of neutrophils, amount of natural killer cells, CD3–/CD19+-cells, CD3+/CD4+-lymphocytes number. Conclusion. Inclusion of immunotropic preparations in complex treatment of secondary pyelonephritis in children is justified in case of detection of above-mentioned disturbances. Key words: secondary pyelonephritis, immunocorrection, hexapeptid, children, azoximer bromide

Quantities and Functions of NK Cells in Severe Aplastic Anemia

IntroductionSevere aplastic anemia (SAA) is an autoimmune disease characterized by bone marrow failure. Natural killer (NK) cells are a key component in the innate immunity and adaptive immune. Our team have previously investigated the quantity and primary functions of NK in SAA. Here, we tend to present the extensive study on the roles NK has played in SAA.MethodsWe successively collected patients with untreated SAA, SAA response to immunosuppressive therapy (SAA-CR+PR), and healthy controls (HC) as research participants. Quantities of circulating NK and their subsets (CD56bright NK and CD56dim NK) were identified, the expressions of surface receptors,intracelluar protein and intracellular cytokines were analyzed by flow cytometry (FCM). Plasmatic IL-2 and IL-12 levels were measured by ELISA. NK cells were cultured with K562 cells. The apoptotic rate of K562 was measured to detect killing functions of NK. NK cells were co-cultured with CD4+ T , CD8+ T and myeloid Dendritic Cells (mDCs) from SAA. Quantities of Th1, Th2, Th17 and Treg, expressions of perforin and Granzyme B in CD8+ T cells and expressions of CD80 and CD86 on mDCs were analyzed by FCM. The mRNA expressions of T-bet, GATA3, RORγt and FOXP3 were measured by qRT-PCR. Proliferation rates of CD8+ T cells and mDC were detected by CCK-8. PeripheralNK cells were sorted and cultured with rhIL-2 and rhIL-12. The supernatants were detected for the cytokines produced by NK. The cytokines and their upstream signal molecules were analyzed by qRT-PCR and Western Blot. Finally, we investigated the quantities and the surface receptors of NK using the mouse model of lymphocyte infusion-induced bone marrow failure to confirm their functions.ResultsThe percentages of NK/lymphocytes and CD56brightNK/NK in SAA were significantly lower than in HC. The absolute number of NK cells in SAA was positively correlated with PLT, absolute number of Reticular cells (Ret) and percentage of Ret (p <0.01). NK cells in SAA expressed higher levels of NKp46, CD158a and perforin compared with those in HC. The percentage of DNAM-1+ NK was higher than that in HC (p=0.007). After PMA stimulation, NK cells in SAA produced lower level of IFN-γ and higher level of IL-10 and TGF-β compared with those in HC. The plasmatic levels of IL-2 and IL-12 in SAA were higher than in SAA-CR+PR and HC. The apoptotic rate of K562 cells when co-cultured with NK cells (1:1) was significantly higher than in HC.IL-2stimulatedNK cells from SAA expressed higher percentage of CD107a+ than from HC (p=0.017) when co-cultured with autologous activated CD4+T cells. After co-cultured with NK cells from SAA, the ratio of Th1/Th2 and Th17/Treg was decreased. The expressions of perforin and Granzyme B in CD8+ T cells were higher when co-cultured with NK from HC than in non-coculture group. The cell inhibition rate of mDCs when co-cultured with NK from SAA was higher than from HC. The expression of CD80 on mDCs when co-cultured with NK from HC was up-regulated.NK cells from SAA secreted lower level of IFN-γ, expressed lower levels of IFN-γ and STAT4 mRNA and phosphorylated STAT4. NK cells from SAA secreted higher levels of IL-10 and TGF-β, expressed higher levels of IL-10, TGF-β and p38 mRNA and phosphorylated p38. Bone marrow failure mouse model showed the quantities of NK cells weredecreased, the expression of NKG2A on NK was elevated. After the treatment of Cyclosporin A (CsA), the quantities NK cells recovered, the expression of NKG2D was elevated while NKG2A was not changed.ConclusionsInsufficient amount of NK cells was shown in SAA patients. The lower quantity of NK cells the patients had,the more severe the disease was. NK cells in SAA were functionally activated and exhibited strong killing abilities. NK cells in SAA produced less Type Ⅰ cytokines and more Type Ⅱ cytokines. Plasmatic levels of IL-2 and IL-12 were elevated in SAA. NK cells from SAA mediated in the differentiation of CD4+ T cells by influencing the balance of Th1/Th2 and Th17/Treg. NK cells from SAA might be able to down-regulate the cytotoxicity activity of CD8+ T cells and the expressions of co-stimulatory molecules on mDCs. The cell signal pathway JAK-STAT4-IFN-γ in NK cells from SAA was down-regulated while pathway p38 MAPK-IL10/TGF-β was up-regulated. Bone marrow failure mouse model suggested the impairment of both quantities and activating status of NK. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

A polymorphic CD4 epitope related to increased susceptibility to Babesia bovis in Canchim calves

Different allelic forms of bovine CD4 were previously described in cattle and were also observed in Canchim calves examined in the present experiment. However, the functional relevance of these different CD4 phenotypes has not yet been investigated. CD4 + T helper cells are known to play a central role in immune control against Babesia bovis infection. Thus, our study aimed to compare the profiles of immune cells, specific antibody titers and blood infection levels measured by qPCR (quantitative polymerase chain reaction) in calves naturally infected with B. bovis, phenotyped as CD4- (absence of anti-CD4 staining), CD4 + (intermediate staining) or CD4 ++ (high staining). The CD4 mRNA precursor was also measured in these animals. Calves with the CD4- phenotype showed higher amounts of B. bovis DNA in blood samples, compared to the other CD4 phenotypes. It was also observed that these calves with higher levels of infection had lower amounts of natural killer cells and higher expression of the CD4 gene, which can be interpreted as a compensation for the failure of the altered CD4 receptor to recognize relevant B. bovis epitopes.

Characterization of natural killer cells in colorectal tumor tissue of rats fed a control diet or a high-fat diet

BackgroundObesity is a major public health problem with an increasing prevalence reaching pandemic levels. The incidence and mortality for colorectal cancer is augmented in overweight and obese individuals. Previous studies demonstrated an impaired number, phenotype and functionality of natural killer (NK) cells under obese conditions. So far, the influence of obesity on NK cells in colorectal cancer tissue remained unclear. Therefore, the aim of the study was to investigate the occurrence and localization of NK cells in colorectal tumors of normal weight and diet-induced obese rats. MethodsWistar rats were fed a normal-fat diet (control) or a high-fat diet (HFD) to induce obesity. In half of the experimental groups azoxymethane (AOM) was injected to induce colorectal cancer. Tumors in colon and rectum were histopathologically classified in adenomas and adenocarcinomas and immunohistologically stained with the rat NK cell marker CD161. Occurrence and localization of NK cells were analyzed and quantified in the tunica mucosa and tunica submucosa of colorectal adenomas and the tunica submucosa of colorectal adenocarcinomas. ResultsNK cells are localized in the tunica mucosa and the tunica submucosa of colorectal tumors with NK cell accumulations as follicle-like aggregates especially in regions of the lamina muscularis mucosae and the lamina propria mucosae of the tunica mucosa as well as in regions of the tunica submucosa adjacent to the lamina muscularis mucosae. Although not statistically significant, the CD161 staining was clearly reduced in the tunica mucosa of colorectal tumors of rats fed a HFD compared to rats fed a control diet. Moreover, the CD161 staining in the tunica mucosa was positively correlated with the final body weight of AOM-treated rats independent of the supplied diet. DiscussionFor the first time, these results provide information about the localization and quantity of NK cells in colorectal tumor tissue of rats fed a control diet or high-fat diet. The slight reduction of NK cell number in colorectal tissue of rats fed a high-fat diet may contribute to an impaired tumor defense and the increased colorectal tumor outcome in diet-induced obese rats.

The immunosuppressive effects of volatile versus intravenous anesthesia combined with epidural analgesia on kidney cancer: a pilot randomized controlled trial.

BackgroundThe aim of this study was to test the hypothesis that the use of inhalational anesthesia leads to higher suppression of the cell-mediated immunity compared to total intravenous anesthesia in patients undergoing kidney cancer surgery under combined low thoracic epidural analgesia and general anesthesia. MethodsPatients were randomly allocated to either propofol-based (intravenous anesthetic) or sevoflurane-based (volatile anesthetic) anesthesia group with 10 patients in each group, along with epidural analgesia in both groups. Amounts of natural killer (NK) cells, total T lymphocytes, and T lymphocyte subpopulations in the blood samples collected from the patients before surgery, at the end of the surgery and postoperative days 1, 3 and 7 were determined by flow cytometric analysis. The NK cell count served as the primary endpoint of the study, whereas the total T lymphocyte count and cell counts for T lymphocyte subpopulations were used as the secondary endpoint.ResultsOur study showed that there were no significant differences in the amount of NK cells, total T lymphocytes, regulatory T cells, and T-helper cells, cytotoxic T lymphocytes, and their subpopulations between the propofol- and sevoflurane-based anesthesia groups when the anesthesia was administered in combination with epidural analgesia.ConclusionsThe results of this pilot study did not support the hypothesis that the use of inhalational anesthesia leads to higher suppression of the cell-mediated immunity than that of total intravenous anesthesia in patients undergoing kidney cancer surgery under combined low thoracic epidural analgesia and general anesthesia.

Fratricide of NK Cells in Daratumumab Therapy for Multiple Myeloma Overcome by Ex Vivo-Expanded Autologous NK Cells.

Purpose: Daratumumab and its use in combination with other agents is becoming a new standard of care for the treatment of multiple myeloma. We mechanistically studied how daratumumab acts on natural killer (NK) cells.Experimental Design: Quantities of NK cells in peripheral blood and/or bone marrow of patients with multiple myeloma or healthy donors were examined by flow cytometry. NK-cell apoptosis and the associated mechanism were assessed by flow cytometry and immunoblotting. Patients' NK cells were expanded in vitro using feeder cells. Combination treatment of daratumumab and expanded NK cells was performed using an MM.1S xenograft animal model.Results: CD38-/low NK cells survived, whereas CD38+ NK cells were almost completely eliminated, in peripheral blood and bone marrow of daratumumab-treated multiple myeloma patients. NK-cell depletion occurred due to daratumumab-induced NK-cell fratricide via antibody-dependent cellular cytotoxicity. Consequently, CD38-/low NK cells were more effective for eradicating multiple myeloma cells than were CD38+ NK cells in the presence of daratumumab. Blockade of CD38 with the F(ab)2 fragments of daratumumab inhibited the antibody-mediated NK-cell fratricide. CD38-/low NK cells displayed a significantly better potential for expansion than CD38+ NK cells, and the expanded NK cells derived from the former population were more cytotoxic than those derived from the latter against multiple myeloma cells. Therefore, infusion of ex vivo-expanded autologous NK cells from daratumumab-treated patients may improve the antibody therapy.Conclusions: We unravel a fratricide mechanism for daratumumab-mediated NK-cell depletion and provide a potential therapeutic strategy to overcome this side effect in daratumumab-treated patients with multiple myeloma. Clin Cancer Res; 24(16); 4006-17. ©2018 AACR.

Transcutaneous Electrical Acupoint Stimulation Improves Immunological Function During the Perioperative Period in Patients With Non-Small Cell Lung Cancer Undergoing Video-Assisted Thoracic Surgical Lobectomy.

The immunological function of patients with malignant tumors may be suppressed during the perioperative period. However, details on the effects of transcutaneous electrical acupoint stimulation (TEAS) on immunological function are relatively lacking. We designed this study to examine the effects of TEAS on the immunological function of patients with non-small cell lung cancer (NSCLC) during the perioperative period. Participants (n = 144) were enrolled and randomly assigned into group TEAS or group sham TEAS. TEAS on bilateral Feishu (BL13), Hegu (L14), and Zusanli (ST36) was performed continuously throughout the procedure. The primary outcome was the quantities of natural killer (NK) cells at 30 minutes before induction (T0), 5 minutes after intubation (T1), at the beginning of the operation (T2), at the beginning of the lobectomy (T3), at the beginning of the lymphadenectomy (T4), and immediately after extubation (T5). The secondary outcomes were the serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) at T0 to T5, the mean arterial pressure (MAP) and heart rate (HR), the intraoperative consumption of propofol and remifentanil, the incidence of hypoxemia, postoperative nausea and vomiting (PONV), and the length of hospital stay. The quantities of NK cells were decreased in group sham TEAS after intubation compared to that in group TEAS, while the quantities of NK cells in group TEAS were similar at T0 to T5. Meanwhile, the quantities of NK cells in group sham TEAS at T1 (P = .012), T2 (P < .001), T3 (P = .027), T4 (P = .045), and T5 (P = .021) were lower than those in group TEAS. In group TEAS, the serum levels of TNF-α were lower at T1 to T5, while the levels of IL-6 were lower at T2 to T5. Furthermore, the intraoperative MAP and HR were more stable, the total propofol and remifentanil consumptions were lower, and the length of hospital stay was shorter than those in group sham TEAS. The application of TEAS can effectively reverse the decrease in NK cells, decrease the serum levels of TNF-α and IL-6, maintain hemodynamic stability during the perioperative period, decrease the consumption of propofol and remifentanil, and shorten the length of the hospital stay.

Programmed differentiated natural killer cells kill leukemia cells by engaging SLAM family receptors.

Natural killer (NK) cells are important innate immune cells that can directly kill transformed or virus-infected cells. The adoptive transfer of NK cells has been used to treat hematological malignancies; however, the limited sources and quantities of NK cells have restricted their clinical application. Here, we acquired sufficient quantities of functional NK cells from CD34+ cells treated with a cytokine cocktail. Microarray analysis of the cultured cells revealed a two-stage pattern of programmed differentiation during NK cell development. Different transcription factors were enriched during these two stages, suggesting that preparation of progenitors committed to the NK cell lineage occurs in program 1, while NK cell transformation and maturation occur in program 2. Cultured NK cells highly expressed signaling lymphocytic activation molecule (SLAM) family receptors (SFRs), while leukemia cells expressed SFR ligands. The engagement of these SFRs strengthened the cytotoxicity of NK cells toward leukemia cells. These results demonstrate a simple method of obtaining sufficient NK cells for clinical application, and have categorized NK cell differentiation according to commitment and transformation programs. Moreover, the binding between SFRs on NK cells and their ligands on leukemia cells suggests a new basis for NK cell therapy for treatment of leukemia.

Immune Cell Repertoire and Their Mediators in Patients with Acute Myocardial Infarction or Stable Angina Pectoris.

Background: To evaluate the natural innate and adaptive immunity through gene expression and cytology levels in peripheral blood mononuclear cells in patients with acute myocardial infarction (AMI), stable angina pectoris (SAP) and controls.Methods: 210 patients with AMI, 210 with SAP, and 250 clinical controls were recruited. Whole human genome microarray analysis was performed in 20 randomly chosen subjects per group were examined to detect the expressions of complement markers, natural killer cells, T cells and B cells. The quantity of these cells and related cytokines as well as immunoglobulin levels were measured in all subjects.Results: In AMI group, the mRNA expressions of late complement component, markers of natural killer cells, CD3+, CD8+ T cells and B cells were down-regulated, while those of early complement component and CD4+T cells were up-regulated (p<0.05). In both AMI and SAP patients, the quantity of natural killer cells, CD3+, CD8+ T cells, B cells, IgM and IgG were significantly lower than those of the controls. CD4+ T cells, CH50, C3, C4, IL-2, IL-4, IL-6 and IFN-γ were significantly higher (p<0.05).Conclusions: In AMI patients, both of gene expressions related to complement, natural killer cells, CD3+, CD8+ T cells, B cells and the quantity of these immune cells decreased while cell number reduced in SAP patients. Immune function in both AMI and SAP patients decreased especially in AMI patients with declined gene and protein levels. To improve the immune system is a potential target for medical interventions and prevention in AMI.

Sea buckthorn (Hippophae rhamnoides L.) oil protects against chronic stress-induced inhibitory function of natural killer cells in rats.

Chronic stress can suppress natural killer (NK) cell activity; this may also be related to the effect of stress on the neuroendocrine-immune network. Sea buckthorn (SBT) (Hippophae rhamnoides L.) is a thorny nitrogen fixing deciduous shrub, native to both Europe and Asia. It has been used as a medicinal plant in Tibetan and Mongolian traditional medicines. SBT has multifarious medical properties, including anti-fatigue as well as immunoregulatory effects. This study reports the effects of SBT oil with regard to the cytotoxicity and quantity of NK cells in the blood of a chronic-stress rat model, in addition to its mechanisms on the neuroendocrine-immune network. These results show that SBT oil, given by gavage to rats with chronic stress, could increase the following: body weight, NK cell quantities, and cytotoxicity, as well as the expression of perforin and granzyme B. The results also show that SBT oil in rats with chronic stress could suppress cortisol, ACTH, IL-1β and TNF-α levels, in addition to increasing 5-HT and IFN-γ serum levels. This leads to suggest that SBT oil, in rats with chronic stress, can increase NK cell cytotoxicity by upregulating the expression of perforin and granzyme B, thus causing associated effects of SBT oil on the neuroendocrine-immune network.

IL-15R -IgG1-Fc Enhances IL-2 and IL-15 Anti-tumor Action through NK and CD8+ T Cells Proliferation and Activation

Natural killer (NK) cells-based immunotherapy is one of the most promising treatments for incurable malignant tumors. NK cells in combination with monoclonal antibodies to surface antigens of the tumor cell have been proved to be effective in a number of clinical trials. A limiting step in the development of successful cellular immunotherapy lies in developing an efficient and economic method to expand appropriate amount of NK cells and CD8(+) T cells. In this study, we constructed a humanized IL-15Ralpha-IgG1-Fc, which mimicked IL-15 trans-presentation. The feasibility of expanding populations of the human NK and CD8(+) T cells by IL-15Ralpha-IgG1-Fc complexes was tested. We then measured the cytotoxicity of expanded NK and CD8(+) T cells against tumor cell lines and primary tumor cells. When tested ex vivo, IL-2/IL-15Ralpha-IgG1-Fc complexes significantly enhanced NK and CD8(+) T cells expansion, isolated or non-isolated from PBMCs. The effect of IL-15Ralpha-IgG1-Fc was dependent on the presence of IL-2 or IL-15. IL-15Ralpha-IgG1-Fc complexes increased NK, CD8(+) T and NKT cells ratio in PBMC and BMMC after 14 days incubation. High level of granzyme B expression was observed in the supernatant of the complexes-treated NK cells. Expanded NK and CD8(+) T cell populations had cytotoxic function against the PC3, LNCaP, K562 and chronic lymphocytic leukemia (CLL) patient primary B cell lymphoma. We concluded that IL-2/IL-15Ralpha-IgG1-Fc significantly enhanced NK, CD8(+) T and NKT cells expansion, which possess strong anti-tumor activity. These data support clinical testing IL-2/IL-15Ralpha-IgG1-Fc expanded NK cells in patients with prostate cancer and CLL.

Differences in natural killer cell quantification and receptor profile expression in HIV-1 infected Chinese children

Natural killer (NK) cells are believed to play a role in the progression of human immunodeficiency virus 1 (HIV-1) disease, and NK cell levels are reduced in individuals with chronic HIV-1 infection. To assess the effects on quantity of NK cells and the changes of NK cell receptors in HIV-1 infected children via mother-to-child transmission, the percentage of NK cells is quantified and the changes in the NK cell receptor profiles in 20 HIV-1 infected children who are not progressing into AIDS were examined. The results showed that NK cell percentage was decreased in the HIV-1 infected children. The expression of NKp30 on NK cells was increased, while the expressions of CD16, NKp44, NKp46, NKp80, NTB-A, CD244, KIR2D, KIR3DL1 and NKG2D on NK cells were decreased in the HIV-1 infected children. NK cell cytolytic activity was elevated in HIV-1 infected children. These results indicate that the acute changes in NK cell percentage and NK cell receptors in HIV-1 infected children are different from the HIV-1 infected adult individuals. Moreover, serum concentrations of IL-18 were elevated in HIV-infected children compared to HIV-uninfected controls. These differences probably play a role in protecting against transmission of maternal HIV-1 virus and guiding the therapeutic strategies for HIV-1 infected children.

Oculocutaneous albinism accompanied by minor morphologic stigmata and reduced number and function of NK cells. A new variant of NK cell defect?

We describe the case of a 7-year-old girl with an apparently new genetic disorder characterized by oculocutaneous albinism, microcephaly, low-set helices, a prominent nose with a broad bridge, a long philtrum, a thin upper lip, a short neck, brachydactyly of the hands and syndactyly between the second and third toes of both feet, thrombocytopenia, and granulocytopenia. In addition, she had extremely low amounts of natural killer cells that were phenotypically normal but lacking cytotoxic activities. Clinically this defect was associated with recurrent and severe respirator-dependent pneumonia of viral and bacterial origin. We assume that the girl presented here represents a similar but distinct entity to the previously described syndromes involving oculocutaneous albinism.

Age-related changes of the function and amount of natural killer cells in the liver: experiment with mice

To investigate the age-related changes of the function and amount of natural killer (NK) cells in the liver. C57BL/6 mice aged 2 weeks and 8 weeks were killed and their livers and spleens were taken out. The percentages and absolute values of the NK cells in the livers and spleens were examined by flow cytometry. Using YAG-1 cells as the target cells, the cytotoxicity of the NK cells was detected. Using thymocytes of the homologous mice as the target cells, the cytotoxicity of the NK T cells was detected. The cytotoxicity of the cytotoxic T lymphocytes (CTLs) was detected with the EL-4 cells as the target cells. RT-PCR was used to detect the mRNA expression of perforin. The NK cell percentage in the liver of the 2-week-old mice was 11.9% +/- 1.7%, not significantly different from that of the 8-week-old mice (9.9% +/- 1.6%, P > 0.05). The NK cell amount of per weight liver tissue of the 2-week-old mice was (11.6 +/- 2.5) x 10(5)/g, significantly higher than that of the 8-week-old mice, (3.4 +/- 0.8) x 10(5)/g, (P < 0.05). The cytotoxicity of the liver NK cells of the 2-week-old mice was 71.0% +/- 5.5%, significantly higher than that of 8-week-old mice (23.8% +/- 4.4%, P < 0.05). The cytotoxicity levels of the NK T cells and CTLs were not significantly different between the mice of different age. The cytotoxicity level of the spleen NK cells was not significantly different between the 2-week-old and 8-week-old mice as well. Phenotypic characterization showed that the NK cells in the liver of the 2-week-old mice were CD69(high) MAC-1(low) LY49C/1(low), whereas those in the liver of the 8-week-old mice were inverse, i.e., CD69(low) MAC-1(high) LY49C/1(high). The level of perforin mRNA was remarkably higher in the lever of the 2-week-old mice than in the liver of the 8-week-old mice. The liver NK cells of the young mice have a higher level of cytotoxicity and a unique phenotype. The liver of young mice has its unique microenvironment and their liver NK cells have a special development pathway.

Distribution pattern of lymphocyte subpopulations in human tonsils as analysed by monoclonal antibodies with double immunoenzymatic labeling

The distribution of B- and T-lymphocytes, T-cell subpopulations, including natural killer cells and monocytes/macrophages, was studied in cryostat sections of human tonsils by the avidin-biotin-peroxidase technique using monoclonal antibodies. A double immunostaining procedure was also developed to detect two types of lymphocytes on one single section simultaneously using horseradish peroxidase and alkaline phosphatase as labeling enzymes. The primary follicles and the germinal centers of the secondary follicles were mainly found to be positive for B-cells. T-cells were predominantly localized in the follicular caps and in the interfollicular areas. The ratio of helper/inducer cells to suppressor/cytotoxic cells was in favour of helper T-cells. Both subpopulations were also predominant in follicular caps and interfollicular areas. The quantity of natural killer cells was very variable and nearly all were localized exclusively in the germinal centers. Monocytes/macrophages were only seen occasionally in the interfollicular areas. The double-immunoenzymatic labeling was useful for the visualization of combinations of antigens, however, without demonstrating the presence of two different surface antigens on one single cell.

Quantitative and functional abnormalities of total T lymphocytes in relatives of patients with Hodgkin's disease

Seven patients, long-term survivors of Hodgkin's disease, and 24 of their relatives (parents, siblings and children), together with normal controls were studied for percentages, absolute counts and mitogen-proliferative responses by means of monoclonal antibodies, E rosette technique and in vitro cultures with PHA, ConA and PWM. The aim of the study was to ascertain whether the impaired cell-mediated immunity of Hodgkin's patients was also present in relatives in order to elucidate the still debated etiology of the defect and of the disease (congenital? environmental? infectious?). The results show that both Hodgkin's patients and their relatives have a significant decrease of total T cells (as T3+, T11+ and E rosette-forming cells) in peripheral blood and a significant impairment of polyclonal responses to all the mitogens employed. The Leu-7+ cells (i.e. a consistent amount of natural killer cells) are significantly increased only in the Hodgkin's patients but not in their relatives. The T cell subpopulations (T4 and T8), B cells and monocytes do not show any difference between the patients, their relatives and normal controls. Our results seem to support, at least in part, the presence of a common defect of T cell lineage both in patients and in their relatives, but its etiology still remains uncertain (genetic? environmental?).