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PubMed1984

The migration of lymphocytes across specialized vascular endothelium: VIII. Physical and chemical conditions influencing the surface morphology of lymphocytes and their ability to enter lymph nodes.

The introductory review amplifies the finding that simply holding lymphocytes in vitro reversibly compromises their ability to enter lymph nodes from the blood, although entry into the spleen is unaffected. The differential migration of T and B lymphocytes from the blood, lymphocyte traffic in athymic rats, and the secretion of a sulphated glycoconjugate by high endothelial cells in lymph nodes are also discussed. Original data are presented concerning the effects of varying the conditions under which lymphocytes are held in vitro (time, temperature, medium, centrifugation) on their ability to enter lymph nodes and also on their surface morphology. In general, conditions that reduced the number of microvilli and induced surface blebbing also tended to affect the delicate function of crossing specialized vascular endothelium; but there was no simple relationship between morphology and migratory behavior. The localization of lymphocytes to the bone marrow was augmented by holding them in vitro, and this effect was greater after holding at room temperature (RT) than at 0 degree C, in contrast to impaired entry into lymph nodes. Small amounts of heparin (10 units) injected along with lymphocytes significantly reduced early localization in lymph nodes. These findings have practical implications for the design of lymphocyte traffic experiments and are relevant to the mechanism of lymphocyte attachment to vascular endothelium, since the well-known effect of trypsinizing lymphocytes can be reproduced by maintenance in vitro.

Animals↗
PubMed1987

Sialylation patterns of lymphocyte function-associated antigen 1 (LFA-1) differ between T and B lymphocytes.

Lymphocyte function-associated antigen 1 (LFA-1) was immunoprecipitated from various types of surface-radioiodinated murine lymphocytes, and analyzed by two-dimensional polyacrylamide gel electrophoresis. LFA-1 alpha and beta chains from splenic B lymphocytes had the same apparent molecular weights as, but distinct isoelectrofocusing patterns from, their counterparts from thymocytes or splenic T lymphocytes. The splenic B lymphocytes lacked a basically charged population of alpha chain, while the thymocytes and the splenic T lymphocytes showed both the acidic and the basic portions. Furthermore, the beta chain of the former migrated more towards the acidic end than that of the latter. No difference was found between LFA-1 molecules of the same lineage of cells from several strains of mice whose H-2 haplotypes were different from one another. When murine lymphocyte lines were examined, LFA-1 with various isoelectrofocusing patterns were recognized. The charge difference again reflected the difference in lymphocyte lineage, but in a more exaggerated manner than that seen with cells from mice. The average acidity of both chains of LFA-1 decreased in the order of B cell lines, pre-B cell lines and T cell lines. The lineage-dependent charge difference of either chain disappeared after neuraminidase treatment of LFA-1, indicating that lymphocyte differentiation was accompanied by changes in LFA-1 sialylation.

Animals↗
PubMed1985

Y+- and L-system amino acid transport in normal and chronic lymphocytic leukemia lymphocytes: photoinhibition by fluoronitrophenylazide.

Three major pathways mediate amino acid transport into mammalian cells: the A-system and the ASC-system, which require a sodium gradient across the plasma membrane, and the L-system, which has no requirement for a sodium gradient. We have found that the lymphocytes from patients with B-cell chronic lymphocytic leukemia (CLL) have a marked reduction in the L-system of amino acid transport when compared to normal human B-lymphocytes from blood or tonsils. Transport by the A- and ASC-systems was not decreased in CLL B-lymphocytes. Because of the specific defect of the sodium-independent L-system amino acid transport in CLL cells, we have examined the activity of another sodium-independent transport system, the Y+-system, in human lymphocytes. The Y+-system favors the transport of dibasic, cationic amino acids such as lysine, ornithine, and arginine, which carry a positively charged group on their side chains. Our studies indicate that there is a large nonsaturable component of amino acid transport by the Y+-system in human lymphocytes. Using a multicomponent mathematical analysis, we have determined that the saturable component of Y+-transport is similar in T- (thymus-derived) and B- (bone-marrow-derived) lymphocytes and is unimpaired in CLL B-lymphocytes. Further, fluoronitrophenylazide, which was thought to be a specific inhibitor of the Y+-system when photoactivated, also inhibits A-, and L-system transport in CLL, T-, and B-lymphocytes.

Amino Acids↗
PubMed1996

The superantigen exfoliative toxin induces cutaneous lymphocyte-associated antigen expression in peripheral human T lymphocytes.

Several immune-mediated dermatoses including psoriasis and atopic dermatitis can be exacerbated by bacterial infections. Superantigen producing bacteria can be isolated from skin lesions of these dermatoses. Consistent with superantigen effects, skewed T cell receptor variable gene usage has been demonstrated within these lesions. Therefore, the question arises whether superantigen induce a skin-seeking phenotype within peripheral T cells. In this study, we investigated the in vitro influence of the V beta 2-selective superantigen exfoliative toxin from Staphylococcus aureus on the expression of the cutaneous lymphocyte-associated antigen on peripheral T lymphocytes of healthy donors. We demonstrate that exfoliative toxin dramatically upregulates cutaneous lymphocyte-associated antigen expression on T cell receptor V beta 2+ lymphocytes. Up to 69% of V beta 2+ lymphocytes expressed cutaneous lymphocyte-associated antigen after 5 days of in vitro culture. Additionally, exfoliative toxin also increased cutaneous lymphocyte-associated antigen expression in CD3+ T cell receptor V beta 2- lymphocytes indicating a different effect as caused by the superantigen-T cell receptor V beta 2 interaction. Our findings suggest influence of bacterial superantigens on T lymphocyte skin homing in vivo.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1987

Inhibitory effects of tunicamycin on the mixed lymphocyte reaction and mitogen-induced lymphocyte blastogenesis.

The role of asparagine(Asn)-linked saccharides on the surface of lymphocytes in cellular interactions was examined by performing studies on the effects of tunicamycin (TM), which inhibits the glycosylation of proteins N-glycosylated at asparagine residues, on the mixed lymphocyte reaction (MLR) and mitogen-induced lymphocyte blastogenesis of human lymphocytes by measuring 3H-thymidine (TdR) incorporation. Responses were expressed as percentages of that of the control (MLR without TM). The lymphocyte blastogenesis in the one- and two-way MLR were, respectively, 43.1% and 48.0% of the control at 0.1 microgram/ml of TM, and 5.5% and 7.2% at 1 microgram/ml of TM. The inhibitory effect of TM on the one-way MLR was shown using TM-pretreated stimulator cells, TM-pretreated responder cells or both. TM blocked lymphocyte blastogenesis in the secondary as well as in the primary MLR. The inhibitory effect of TM on the two-way MLR was observed when TM was added on day 0 to day 2, but not on day 4 of incubation. TM blocked mitogen-induced lymphocyte blastogenesis by phytohemagglutinin, concanavalin A or by pokeweed mitogen. As TM had no cytotoxic effect on cultured cells, these inhibitory effects of TM were thought to be due to the loss of Asn-linked saccharides from glycoprotein of the surface of lymphocytes. These findings indicated that Asn-linked saccharides of glycoprotein on the surface of lymphocytes were important in cellular interactions that are necessary for the cellular immune response.

Antigens, Surface↗
PubMed1999

A comparison of the expression of lymphocyte activation markers in blood, bronchial biopsies and bronchoalveolar lavage: evidence for an enrichment of activated T lymphocytes in the bronchoalveolar space.

In this study healthy never-smoking subjects (n = 18) were recruited from a population study. Bronchoalveolar lavage (BAL), blood lymphocytes and bronchial biopsies, analysed both in the epithelium and lamina propria, were stained for T and B lymphocytes, natural killer (NK) cells and different subpopulations of T lymphocytes. In BAL, significantly higher proportions of T lymphocytes (CD3), T lymphocyte activation markers; HLA-DR, CD26+, CD49a+, CD54+ and CD69+, helper T (CD3+4+) and memory helper T lymphocytes (CD4+45RO+29+) and memory T lymphocytes (CD3+45RO+) were found, compared to blood. However, the proportion of IL-2 receptor-positive T lymphocytes (CD25+) was lower in BAL than in blood. A previously described higher ratio of CD3+4+/CD3+8+ in BAL than in blood (3.4 vs 1.7; P = 0.001) was confirmed. In bronchial biopsies, we found significantly higher numbers of CD8+ cell profiles per mm2 in the epithelial compared to the lamina propria compartment. We conclude that healthy never-smoking men have higher levels of activated memory T lymphocytes in BAL than in blood, and that the T-cell subpopulations differ in the epithelial compared to the lamina propria compartment in the bronchial mucosa and these compartments should be analysed separately. It is reasonable to think that there is a gradient from blood to the airway lumen where T cells are recruited from blood to take part in the defense towards damaging agents.

Antigens, CD↗
PubMed1987

Lymphocyte subsets in tumour of patients with undifferentiated nasopharyngeal carcinoma: presence of lymphocytes with the phenotype of activated T cells.

We have analyzed lymphocytes infiltrating nasopharyngeal carcinomas, using a combination of immunoperoxidase staining of frozen and paraffin-embedded sections, and immunofluorescence on lymphocyte suspensions recovered from teased tumours. A panel of monoclonal antibodies was used to define lymphocytic subsets on frozen sections of 14 different tumours. The vast majority of peri- and intra-tumoral lymphocytes were stained by OKT3 antibody. In 8 sections, T4 positive cells were largely predominant, while T8 positive cells were the majority in three sections. Twenty-nine paraffin-embedded sections from other NPC patients stained with HNK-1 antibody showed a variable percentage of positive cells reaching 6 to 15% in nine patients. Most HNK-1 positive cells had the morphology of large granular lymphocytes typical of natural killer cells. Double staining experiments on lymphocytes isolated from 7 tumours revealed a constant presence of T3 positive, HLA-DR positive lymphocytes (from 6 to 29% of mononuclear cells), and of lymphocytes coexpressing the T3 and the Tac (IL-2 receptor) antigens (from 5 to 12% of mononuclear cells). Lymphocytes with a phenotype of activated T-cells are thus constantly found in NPC tumours.

Antigens, Surface↗
PubMed1996

Decreased number of CD73 (ecto-5'-nucleotidase) molecules on lymphocytes from patients with primary immunoglobulin deficiencies. Correlation between number of CD73 molecules and T-lymphocyte function in vitro.

CD73 is a bifunctional glycosyl phosphatidylinositol anchored leucocyte differentiation antigen which has specific ecto-5'-nucleotidase (ecto-5'-NT) activity and is an accessory T-lymphocyte activation molecule. The aim of the present study was to investigate the CD73 expression on blood mononuclear cells (BMC) from a group of patients with primary immunoglobulin deficiency (IGD). This group of patients had both significantly decreased levels of ecto-5'-NT on BMC (P = 0.002) and decreased numbers of CD73 molecules per CD73+ lymphocyte (P = 0.01). Five of the 10 patients had a decreased percentage of CD73+ lymphocytes. Among B-lymphocytes the patients had normal percentages of CD73+ cells but four of the 10 patients had numbers of CD73 molecules per CD73+ B-lymphocyte below the normal range. Among CD4-lymphocytes three out of 10 patients had percentages of CD73+ below the normal range and four out of 10 patients had decreased percentages of CD73+ CD8-lymphocytes. Significant correlations were found between in vitro proliferative responses to mitogens and the number of CD73 molecules per CD73+ lymphocyte (rs = 0.60, P < 0.01) and per CD73+ CD8-lymphocyte (rs = 0.64, P < 0.02). In addition, a positive correlation was found between ability to proliferate and level of ecto-5'-NT on BMC (rs = 0.53, P < 0.05). Furthermore the ability of BMC to synthesize ecto-5'-NT was studied. During 2 days culture ecto-5'-NT activity increased markedly on BMC from both patients and healthy donors. The level of activity on BMC from all patients attained levels higher than on freshly isolated BMC from healthy donors. This shows that the decreased levels of ecto-5'-NT found on freshly isolated BMC from patients with IGD is due to defective regulation of the enzyme activity in vivo.

5'-Nucleotidase↗
PubMedDate not supplied

Decreases of natural killer cells and T-lymphocyte subpopulations and increases of B lymphocytes following a 5-day occupational exposure to mixed organic solvents.

The authors examined the effects of organic solvents on lymphocyte subpopulations in blood. Natural killer and T-lymphocyte subpopulations and B (CD19+) -lymphocytes were measured with flow cytometry in 16 male rotogravure printers on a Friday and on the following Monday. Numbers of all 3 subpopulations of natural killer cells (i.e., CD57+ CD16+, CD57- CD16+, and CD57+ CD16- cells), 2 subpopulations of T lymphocytes (CD4+ CD45RA+ and total CD8+ cells), and total lymphocytes on Friday were significantly fewer than those found on the following Monday. Conversely, the number of B lymphocytes on Friday was significantly larger than the number on Monday. The number of B lymphocytes was significantly correlated with blood toluene levels on Friday. The alteration in the number of CD57+ CD16+ NK cells from Friday to the following Monday was correlated inversely with the corresponding change in exposure level of toluene on Friday. The authors suggest that the effects of mixed organic solvents (primarily toluene) are recoverable decreases of natural killer cells and T lymphocytes and increases in B lymphocytes.

Adult↗
PubMed1979

Changes in mixed lymphocyte culture-reactive lymphocytes following alloimmunization of single lymph nodes in sheep.

When an "isolated" single lymph node is challenged with irradiated allogeneic lymphocytes, there is a change in the reactivity of the lymphocytes flowing out of the node when they are cultured in vitro in unidirectional mixed lymphocyte cultures (MLC) against the immunizing lymphocytes. These changes in the reactivity of the recipient lymphocytes are shortlived, follow a set time sequence in relation to the cell traffic changes accompanying the immune response, are the property of small lymphocytes and not blast cells, are exhibited by surface Ig-negative cells, and they are specific for the donor lymphocytes. It is suggested that antigen causes the selective retention of antigen-specific lymphocytes within the stimulated node followed by the proliferation and differentiation of large numbers of antigen-specific cells, which then leave the lymph node as small lymphocytes.

Animals↗
PubMed1980

Studies of lymphocyte proliferation in hairy cell leukaemia: activity in mixed lymphocyte reaction and responses to mitogens.

Subpopulations of splenic lymphocytes from patients with hair cell leukaemia (HCL) were compared with similar subpopulations of lymphocytes from reference individuals for their ability to respond to mitogens and to participate in allogenic and autologous mixed lymphocyte reactions. T cell enriched subpopulations were obtained by double passage of mononuclear cells through mylon wool columns. Non-T cell subpopulations were collected by eluting adherent cells from nylon wool columns and by incubating them with sheep erythrocytes followed by density gradient centrifugation. Unfractionated mononuclear cells, T enriched and non-T subpopulations were compared. Enriched T cell subpopulations from HCL and reference patients responded similarly to allogeneic antigens and phytohaemagglutinin. Splenic non-T cells from reference patients produced a stronger stimulus in the allogeneic mixed lymphocyte reaction than did the unfractionated or the T enriched cells. In contrast, the non-T subpopulations from patients with HCL produced a reduced response compared to that of reference splenic cells when mixed with allogeneic lymphocytes. In addition, non-T cells from HCL patients failed to respond to pokeweed mitogen. Neither reference nor HCL splenic cells produced a significant response in the autologous mixed lymphocyte reactions. The data suggest that splenic non-T cells from patients with HCL either suppress the stimulatory capacity of normal B lymphocytes or fail to stimulate allogeneic lymphocytes in the mixed lymphocyte reactions.

Humans↗
PubMed2006

Platelet-lymphocyte conjugation differs between lymphocyte subpopulations.

BACKGROUND: Platelets can bind to, and thereby influence, lymphocyte function. OBJECTIVE: The propensities of different lymphocyte subpopulations to form platelet-lymphocyte conjugates/aggregates (P-Lym) was investigated using four-color whole blood flow cytometry. RESULTS: P-Lym constituted approximately 3% of circulating lymphocytes. Platelet conjugation was most common among large (monocyte-sized) lymphocytes. Platelet activation by ADP slightly increased platelet-T-cell conjugation, mainly to T-cytolytic (Tc) cells, but markedly elevated platelet-natural killer (NK)-cell conjugation. T-cell activation by phytohemagglutinin increased heterotypic conjugation among both T-helper (TH) and Tc cells, whilst NK-cell activation by interleukin-2 affected platelet-NK-cell aggregation little. Neither platelet activation nor lipopolysaccharides-induced B-cell activation enhanced platelet-B-cell aggregation. Activation-dependent heterotypic conjugation was mainly found among large cells, with increased percentages of conjugated cells and more platelets bound per lymphocyte. P-Lym formation initiated by platelet activation was abolished by P-selectin blockade, and tended to be reduced by inhibition of GPIIb/IIIa, CD11b, or CD40L. P-Lym formation initiated by lymphocyte activation was partially inhibited by each of these blocking agents, but more markedly inhibited when the blocking agents were combined. CONCLUSIONS: Platelets selectively bind to larger and activated lymphocytes. T-lymphocyte activation enhances platelet-T-cell aggregation. Platelet activation enhances platelet-Tc aggregation slightly and platelet-NK-cell aggregation markedly, while cellular activation affects platelet-B-cell aggregation little. P-selectin ligation is essential, but GPIIb/IIIa, CD40L, and CD11b also contribute to the heterotypic conjugation.

Adult↗
PubMed1992

Lymphocyte and lymphocyte subset numbers in blood and in bronchoalveolar lavage and pleural fluid in various forms of human pulmonary tuberculosis at presentation and during recovery.

BACKGROUND: Lymphocytes have a central role in human defences against mycobacteria. A study was designed to assess the relation between lymphocyte responses and clinical pattern of disease, nutrition and recovery during treatment in patients with tuberculosis. METHODS: Lymphocyte numbers and subsets (on the basis of CD3, CD4, and CD8 monoclonal antibodies) were measured in peripheral blood and, where appropriate, bronchoalveolar lavage or pleural fluid of patients with different forms of pulmonary tuberculosis. Eleven had localised pulmonary tuberculosis, 18 miliary tuberculosis and seven a tuberculous pleural effusion. RESULTS: CD4 lymphocytes were found in greatly increased numbers in pleural fluid and were relatively depleted in the blood. Lymphocyte numbers in bronchoalveolar lavage fluid varied widely in localised pulmonary and miliary tuberculosis but were highest in lavage fluid from patients with miliary tuberculosis. This was due to an increase in CD8 lymphocytes, which were also increased in the blood. Lymphocyte numbers bore no relation to nutrition, symptom duration, or radiographic profusion scores. In miliary tuberculosis the time taken for the chest radiograph to clear (mean (SD) 17.6 (7.8) weeks) correlated with lymphocyte numbers in lavage fluid, especially CD8 cells (r = 0.74), but not with the patients' age or nutrition. After 8 weeks' treatment, total and CD4 lymphocyte numbers in lavage fluid showed a substantial increase. CONCLUSION: The association of CD8 cells with delayed recovery is compatible with suppression of the antimycobacterial action of macrophages. The switch to predominance of CD4 cells in lavage fluid during successful treatment supports the view that they may have a role in eliminating mycobacteria.

Adult↗
PubMed1976

Lymphocyte mobilization in response to physical work in sarcoidosis. Influence on lymphocyte subpopulations, DNA synthesis and cytotoxicity.

Peripheral blood lymphocytes of nine patients with pulmonary sarcoidosis were collected before and after a standardized bicycle ergometer test. The lymphocytes were characterized by various cell surface markers as T or B lymphocytes, and functionally by mitogen- or antigen-induced DNA synthesis and by antibody dependent cell-mediated cytotoxicity. Physical work resulted in an increase of circulating lymphocytes. Proportionately more B lymphocytes were mobilized. The DNA synthesis of lymphocytes in response to mitogens and antigen was reduced, whereas the cytotoxicity was augmented. Compared with normals the mobilization of lymphocytes was less, and a deficiency of both T and B lymphocytes was revealed. The functional impairment of lymphocytes in the patient group persisted.

Adult↗
PubMed1989

Lymphocyte membrane alpha-1-acid glycoprotein: a cellular synthesis during lymphocyte activation.

Soluble alpha 1 acid-glycoprotein is considered an "acute phase protein" with an inhibitory effect on lymphocyte activity; it has recently been shown that a lymphocyte modulatory variant of alpha 1 acid-glycoprotein has a positive role on T cell activation. It is not clear whether the presence of this glycoprotein on lymphocyte membranes is due to an endogenous production or to a passive uptake of soluble alpha 1 acid-glycoprotein by its carbohydrate moiety. Our data show an increase of membrane alpha 1 acid-glycoprotein both in peripheral blood lymphocyte and T-enriched lymphocytes after phytohemagglutinin stimulation. Peripheral blood lymphocyte enzymatic treatment by neuraminidase does not affect alpha 1 acid-glycoprotein expression while pronase digestion induces a strong decrease of alpha 1 acid-glycoprotein positive lymphocytes and a resynthesis after phytohemagglutinin stimulation. Furthermore, the presence of alpha 1 acid-glycoprotein was prevalently, found on helper/inducer lymphocytes. These data support the hypothesis of a synthesis of alpha 1 acid-glycoprotein by T lymphocytes during their activation process.

Cell Membrane↗
PubMed1975

B lymphocytes, T lymphocytes and phytohaemagglutinin responsiveness in atopic dermatitis.

The spontaneous binding of sheep erythrocytes to human lymphocytes with the formation of rosettes was used as a measure of thymus-derived lymphocytes and the rosette formation with complement-coated sheep erythrocytes as a measure of bursa-dependent lymphocytes in the peripheral blood of patients with atopic dermatitis. Both percentage proportion and the absolute number of thymus-derived lymphocytes were slightly but significantly reduced. In a few patients with atopic dermatitis the percentage and absolute number of bursa-dependent lymphocytes were increased. However, when all values were considered, no significant differences were demonstrated. The responsiveness of cultured lymphocytes to phytohaemagglutinin was normal when the culture medium contained foetal calf serum. The response of normal lymphocytes was not impaired by serum from patients with atopic dermatitis. However, the response was depressed when atopic dermatitis lymphocytes were cultured in medium containing autologous serum, indication that a partial lymphocyte defect becomes manifest only in the presence of a serum factor, presumably IgE.

Adolescent↗
PubMed1976

Lymphocyte transformation induced by autologous cells. III. Lymphoblast-induced lymphocyte to stimulation does not correlate with EB viral antigen expression or immunity.

Human mitogen-induced and cell line B lymphoblasts stimulate the proliferation of allogeneic and autologous lymphocytes in culture. The role in thes reaction of EB viral determinants on the stimulating cells and immunity of the lymphocyte donor to the EB virus has been studied. The stimulatory capacity of cultured cell line lymphoblasts is not inhibited by incubating lymphoblasts with antisera to EB viral determinants. Cultured cell line B lymphoblasts stimulate as much thymidine incorporation by lymphocytes from donors with or without immunity to the EB virus. Further, a B lymphoblast cell line (U-698) which lacks the EB viral genome stimulated as much lymphocyte proliferation as did B lymphoblasts with the EB genome. Cultured T lymphoblast cell lines do not stimulate allogeneic lymphocyte proliferation. These cells appear to lack the determinants which stimulate lymphocyte transformation. No evidence was found that cultured cell line T lymphoblasts suppressed allogeneic lymphocyte proliferation. Mitogeninduced lymphoblasts from EB-immune and non-immune subjects stimulated the proliferation of autologous lymphocytes comparably. It is concluded that neither immunity to the EB virus nor expression of EB viral antigens on mitogen-induced on cell line lymphoblasts is necessary for the stimulation of lymphocyte proliferation.

Adult↗
PubMed1981

Human T-lymphocyte colonies: generation of colonies in different lymphocyte subpopulations.

The generation of human T-lymphocyte colonies from different lymphocyte subpopulations in the presence of PHA alone, or PHA plus media conditioned by PHA-stimulated lymphocytes (PHA-LCM) has been investigated. The separation technique consisted of phagocytic cell depletion by carbonyl iron treatment and fractionation of non-phagocytic cells (NP cells) into B cells and T + null cells by affinity chromatography on an anti-F(ab')2 column. The T cells were separated from the null cells by E-rosette sedimentation. Under these conditions, we showed that: no T-lymphocyte colonies were obtained from the null-cell subset in the presence of PHA of PHA + PHA-LCM; T-lymphocyte-colony formation potential was retained in the T-cell subset. Some variability was observed in the production of T colonies using peripheral blood lymphocytes (PBL) from different donors. Low producers and high producers of T-lymphocyte colonies were encountered. The low production of T-lymphocyte colonies observed in some donors was due to a suppressive effect mediated by the phagocytic cells, probably monocytes. The anti-F(ab')2 immunoadsorbent retained a cell population necessary for T-lymphocyte colony growth.

Adult↗