Acid Α-naphthyl Acetate Esterase Research Articles

Dual-factor Synergistically Activated ESIPT-based Probe: Differential Fluorescence Signals to Simultaneously Detect α-Naphthyl Acetate and Acid α-Naphthyl Acetate Esterase.

α-Naphthyl acetate esterase (α-NAE) and acid α-naphthyl acetate esterase (ANAE), a class of special esterases, are important for lymphocyte typing and immunocompetence-monitoring. As such, the simultaneous detection of α-NAE and ANAE has become a target to effectively improve the accuracy in lymphocyte typing. Therefore, we developed a dual-factor synergistically activated ESIPT-based probe (HBT-NA) to detect α-NAE and ANAE sensitively, rapidly, and simultaneously in a differential manner. HBT-NA exhibits differential fluorescence signal outputs toward small changes of α-NAE and ANAE activities. HBT-NA displays a weak fluorescence signal at 392 nm over a pH range from 6.0 to 7.4. However, when it interacts with α-NAE (0–25 U) at pH = 7.4, the fluorescence intensity at 392 nm enhanced linearly within 60 s (F392 nm/F0392 nm = 0.042 Cα-NAE + 1.1, R2 = 0.99). Furthermore, HBT-NA emits ratiometric fluorescence signals (F505 nm/F392 nm) for ANAE (0–25 U) at pH = 6.0 within 2.0 min, exhibiting a good linear relationship (F505 nm/F392 nm = 0.83CANAE – 1.75, R2 = 0.99). The differential fluorescence signals can be used to simultaneously detect the activities of α-NAE and ANAE in solutions and complex living organisms. More importantly, based on the differential fluorescence signals toward α-NAE and ANAE, T lymphocytes and B lymphocytes could be successfully typed and differentiated among nontyped lymphocytes, facilitating the real-time evaluation of their immune functions using flow cytometry. Hence, HBT-NA could be used for the ultrasensitive detection of the enzyme activities of α-NAE and ANAE, the real-time precise typing of lymphocytes, and the monitoring of immunocompetence.

Effects of stale maize on growth performance, immunity, intestinal morphology and antioxidant capacity in broilers.

ObjectiveThis study was conducted to determine the effects of stale maize on growth performance, immunity, intestinal morphology, and antioxidant capacity in broilers.MethodsA total of 800 one-day-old male Arbor Acres broilers (45.4±0.5 g) were blocked based on body weight, and then allocated randomly to 2 treatments with 20 cages per treatment and 20 broilers per cage in this 6-week experiment. Dietary treatments included a basal diet and diets with 100% of control maize replaced by stale maize.ResultsThe content of fat acidity value was higher (p<0.05) while the starch, activities of catalase and peroxidase were lower (p<0.05) than the control maize. Feeding stale maize diets reduced (p<0.05) average daily feed intake (ADFI) throughout the experiment, feed conversion ratio (FCR) during d 0 to 21 and the whole experiment as well as relative weight of liver, spleen, bursa of Fabricius and thymus (p<0.05) on d 21. Feeding stale maize diets decreased jejunum villus height (VH) and VH/crypt depth (CD) (p<0.05) on d 21 and 42 as well as ileum VH/CD on d 42. The levels of immunoglobulin G, acid α-naphthylacetate esterase positive ratios and lymphocyte proliferation on d 21 and 42 as well as lysozyme activity and avian influenza antibody H5N1 titer on d 21 decreased (p<0.05) by the stale maize. Feeding stale maize diets reduced (p<0.05) serum interferon-γ, tumor necrosis factor-α, interleukin-2 on d 21 and interleukin-6 on d 21 and 42. Broilers fed stale maize diets had lower levels of (p<0.05) total antioxidative capacity on d 42, superoxide dismutase and glutathione peroxidase on d 21 and 42, but higher (p<0.05) levels of malondialdehyde on d 21 and 42.ConclusionFeeding 100% stale maize decreased ADFI and FCR, caused adverse effects on immunity and antioxidant function and altered intestinal morphology in broilers.

Enantioselective Interaction of Acid α-Naphthyl Acetate Esterase with Chiral Organophosphorus Insecticides

Many previous works have demonstrated that acetylcholinesterase (AChE) was enantioselectively inhibited by chiral organophosphorus insecticides (OPs) and that a significant difference in reactivation existed for AChE inactivated by (1R)- versus (1S,3S)-stereoisomers of isomalathion. It had been known that α-naphthyl acetate esterase (ANAE), an enzyme which might play an essential role in the growth of plants and the defense of plants against environmental stress by regulating the concentration of hormones in plants, can be inhibited by OPs. However, it was unknown whether interaction of ANAE with chiral OPs was enantioselective. The present work investigated the inhibition kinetics and spontaneous reactivation of ANAE inactivated by enantiomers of malaoxon, isomalathion, and methamidophos. The order of inhibition potency is (R) > (S) for malaoxon, (1R,3R) > (1R,3S) > (1S,3R) > (1S,3S) for isomalathion, and (S) > (R) for methamidophos according to bimolecular rate constants of inhibition (ki), which is consistent with the order observed in the enantioselective inhibition of AChE by malaoxon, isomalathion, and methamidophos. The difference in spontaneous reactivation of AChE inactivated between (1R)- and (1S,3S)-isomers of isomalathion is conserved for ANAE. The observations indicated ANAE and AChE have similar selective inhibition kinetics and postinhibitory reactions in reaction with chiral OPs.

Effects of vitamin E and selenium yeast on growth performance and immune function in ducks fed maize naturally contaminated with aflatoxin B1

This study was conducted to determine the effects of a combined vitamin E (VE) and selenium yeast (SeY) supplementation on growth performance, blood profiles, relative organ weight, and immune function in ducks fed diets containing maize naturally contaminated with a variety of mycotoxins, but primarily aflatoxin (AFB1). A total of 448 ducks were blocked on the basis of sex and body weight, and then randomly assigned to 1 of 4 treatments in a 2×2 factorial arrangement with 2 levels of AFB1 contaminated maize (0 or 100%) and 2 VE and SeY levels (0IU VE and 0mgSeY/kg or 50 IU VE and 0.2mgSeY/kg) for a 28-d study period. There were 7 replications (pens) per treatment and 16 birds per pen. Detectable levels of other toxins were present but that only AFB1 exceeded limits, and the level of the AFB1 was 196.8μg/kg in the contaminated maize, 130.5 and 142.4μg/kg in the 100% contaminated starter diets with or without VE and SeY, respectively, and 152.6 and 157.1μg/kg in the 100% contaminated grower diets with or without VE and SeY, respectively. Ducks fed AFB1 contaminated maize diets had lower (P<0.05) ADG and ADFI, and greater (P<0.05) death rates during the whole experiment. The supplementation of VE and SeY increased (P<0.05) FCR during d 0 to 14 but decreased (P<0.05) ADFI during d 15 to 28 and 0 to 28. Interactions were observed (P<0.05) in ducks fed AFB1 contaminated diets. Feeding AFB1 contaminated maize diets increased (P<0.05) the activity of lactate dehydrogenase, whereas the levels of total protein, albumin, and globulin decreased (P<0.05) at both d 14 and 28. However, VE and SeY had no consistent effects. The lymphocyte proliferation and acid α-naphthylacetate esterase positive ratios (ANAE+) were decreased (P<0.05) by the inclusion of AFB1 contaminated diets during the whole experiment but benefited from VE and SeY supplementation. Dietary VE and SeY improved (P<0.05) the levels of interleukin 2 (IL-2) and interleukin 6 (IL-6) throughout the experiment. Avian influenza (H5N1) antibody titer was reduced (P<0.05) by the inclusion of AFB1 contaminated maize without VE and SeY supplementation. Ducks fed AFB1 contaminated maize diets had greater (P<0.05) relative weights of spleen and bursa of fabricius but lower (P<0.05) thymus, and the supplementation of VE and SeY increased (P<0.05) the relative bursa of fabricius weight in the presence of contaminated maize. Interactive effects were observed (P<0.05) in ANAE+ and IL-2 on d 28 and relative weight of bursa of fabricius. In conclusion, feeding AFB1 contaminated maize caused adverse effects on growth performance, immunity, and relative immune organ weight in ducks, while dietary addition of VE and SeY partially counteracted the negative effects of AFB1 on growth and death rate.

Stereoselective toxicity of malathion and its metabolites, malaoxon and isomalathion

In recent years, the stereoselective toxicity of chiral organophosphorus pesticides has received increasing attention from environmental toxicology scientists. In an attempt to explore the stereoselective effects of chiral organophosphorus pesticides on non-target organisms and enzymes, this work investigated the stereoselective toxicity and inhibition of malathion, malaoxon and isomalathion on Daphnia magna and on acid α-naphthyl acetate esterase extracted from wheat flour, respectively. Toxicity differences were observed between the two enantiomers of malathion and malaoxon and among the four stereoisomers of isomalathion in the LC50 values on D. magna and the IC50 values on acid α-naphthyl acetate esterase. Enantiomers with (R)-configuration in all the tested compounds showed higher toxicity to D. magna than their (S)-forms and racemic forms. Racemic malaoxon and isomalathion showed the strongest inhibition on acid α-naphthyl acetate esterase compared to their respective enantiomers and stereoisomers.

Polysaccharide isolated from Glycyrrhiza uralensis Fisch induces intracellular enzyme activity of macrophages

A purified fraction of water-soluble polysaccharides was isolated from Glycyrrhiza uralensis Fisch using ion exchange and size exclusion chromatography. The present study was undertaken to discuss the preliminary immunoregulation mechanism of glycyrrhiza polysaccharide (GP) by cytochemistry and quantitative analyses, and intracellular enzyme measurement of macrophages. Acid phosphatase (ACPase), adenosine triphosphatase (ATPase), acid α-naphthyl acetate esterase (ANAE) and succinate dehydrogenase (SDH) in macrophages were stained with different methods. The results indicate that GP increased the production of ACPase, ATPase, ANAE and SDH; the activities of lysozyme (LSZ) and superoxide dismutase (SOD) of macrophages were also induced by GP. Our data suggest that the beneficial therapeutic effects of GP may be attributed partly to its ability to modulate macrophage immune functions.

Determination of acid α-naphthyl acetate esterase enzyme activity in peripheral blood leukocytes of gazelles (Gazella subgutturosa)

We examined gazelle peripheral blood leucocytes using the α-Naphthyl acetate esterase (ANAE) staining technique (pH 5.8). Our purpose was to determine the percentage of ANAE positive lymphocytes. The proportion of ANAE positive T-lymphocytes was 72%. T-lymphocytes showed an ANAE positive reaction, but eosinophilic granulocytes and monocytes also showed a positive reaction. By contrast, no reaction was detected in B-lymphocytes, neutrophil granulocytes or platelets. The reaction observed in T-lymphocytes was a red-brown coloration, usually 1–2 granules, but enough granules to fill the cytoplasm were detected rarely. As a result of ANAE enzyme staining, we concluded that the staining technique can be used as a cytochemical marker for gazelle T-lymphocytes.

Classification of haematopoietic cells and haemocytes in Chinese prawn Fenneropenaeus chinensis

It is commonly believed that crustacean haemocytes originate from a specialised haematopoietic tissue (HPT), whereas the differentiation relationship between HPT cells and circulating haemocytes is still not clearly understood. The HPT cells and haemocytes of Fenneropenaeus chinensis were characterised using morphological and histochemical methods. Three types of HPT cells were identified under the transmission electron microscope (TEM). Type 1 cells had high N/C ratios, developed dispersed chromatins and no cytoplasmic granules. Type 2 cells had smaller size, developed condensed chromatins and cytoplasmic granules, which were homogeneous or striated in type 2a cells, and homogeneous in type 2b cells. We deduce that type 1 cells may give rise to type 2 cells in terms of the presence of possible intermediates between type 1 and type 2 cells. The circulating haemocytes were divided into three populations, i.e. hyaline haemocytes (HH), small granular haemocytes (SHG) and large granular haemocytes (LGH), based on Wright–Giemsa staining and TEM observation. Comparing the HPT cells with the circulating haemocytes, type 2a cells of HPT may represent the HH due to similar granule types, cell size and N/C ratios, and type 2b cells may be the young and immature LGH. By Wright–Giemsa and acid α-naphthyl acetate esterase staining, the intermediates between the HH and SGH were observed, which indicates that the SGH may be derived from the HH in the circulatory system. Therefore, it is suggested that the F. chinensis haemocytes could be divided into two haemocyte lineages, i.e. the HH-SGH and LGH lineage.

Quantitative and functional analysis of peripheral blood mononuclear cells in European brown bear ( Ursus arctos arctos)

Methodology was established to study relative amounts of mononuclear white blood cell populations (small lymphocytes and monocytes) and lymphocyte functions at different seasons in European brown bear ( Ursus arctos arctos). May-Grünwald-Giemsa (MGG) staining was used for morphological differentiation of mononuclear cell populations. Monocytic and T lymphocytic populations were identified using enzymatic ANAE (acid α-naphthyl acetate esterase) marker. T lymphocytes were also characterized by T-cell rosetting with sheep red blood cells (SRBC rosetting method). Lymphocyte functions were studied by measuring the proliferative reactivity of these cells in response to mitogenic (PHA) and antigenic (PPD) stimulus. Both the rosetting method and the ANAE marker were found suitable for identifying bear MNL populations as long as no mononuclear antibodies are available for these cells. However, the ANAE method was considered more convenient because it allows identification of a larger number of cell populations. The MGG staining was found useful as a standard method by which the successful staining of ANAE preparations could be controlled. The relative number of monocytes was found decreased during the denning period. No seasonal variations were observed in the relative number of small lymphocytes or in the ratio of T to B cells. T-cell reactivity against a mitogenic stimulus appeared steady all through the year, whereas proliferative response against antigen stimulation was found reduced during the February denning period and in March.

Immunosuppression in broilers under experimental aflatoxicosis

The effects of purified aflatoxin B 1 on cell mediated immunity in broilers were carried out using 0·3 p.p.m. and 1 p.p.m. dose level. The percentage of acid α-naphthyl acetate esterase reacting lymphocytes was significantly ( P < 0·05) decreased in chicks fed 0·3 p.p.m. and 1 p.p.m. level of aflatoxin B 1. The albumin and globulin values were also significantly decreased ( P < 0·05) at both dose levels.

Variation in the composition of gingival inflammatory cell infiltrates

Biopsy specimens were taken at gingivectomy from 18 adult patients undergoing treatment for chronic marginal periodontitis. They were embedded so that the cut surface of the gingiva was parallel to the top of the block to obtain a comprehensive view in a transversal plane of the inflammatory cell infiltrate near the bottom of the pocket. Sections were stained with HES or with toluidine blue for histological description, and acid alpha-naphthyl acetate esterase (ANAE) was used to differentially stain T lymphocytes, plasma cells and monocytes/macrophages. Sections stained with HES showed that the density and size of the cell infiltrates varied along the circumference of a tooth over very short distances and on various surfaces on neighbouring teeth. Differential counts of cells stained for ANAE demonstrated great variation in the composition of the cell infiltrates, particularly along the pocket epithelium. The predominating ANAE positive cell type in this area was T lymphocytes, while in the central connective tissue, plasma cells predominated. There was no systematic covariation between the localization of the gingiva (i.e. mesial, facial, etc.) and the composition of the cell infiltrates. The local variation in the composition of the cellular infiltrate most likely reflects local variability in the noxious substances (i.e. plaque composition) within the periodontal pocket, and in the resulting local inflammatory response.

Acid .ALPHA.-naphthyl acetate esterase(ANAE) staining of porcine lymphocytes.

Acid α-naphthyl acetate esterase (ANAE) activity was investigated to be one of porcine T lymphocyte markers by modified fixation procedures in normal porcine lymphocytes prepared from spleen, blood, mesenteric lymph nodes, thymus and Peyer's patch. Smears were fixed for 2 to 3 seconds with buffered formalin acetate (pH 6.6). The majority of ANAE positive cells had the red-brown spot. The cells diffusely stained with densely red color were found to be monocytes. Other stained cells had stained granules. There were no differences between the ratios of ANAE positive cells with stained spots or granules and those of E-rosette positive cells in lymphocyte preparations from spleen, blood, mesenteric lymph nodes and Peyer's patch. However, the ANAE positive cells in thymus were stained faintly and the ratio of ANAE positive cells was by far lower than that of E-rosette positive cells. In Con A or PHA-P stimulated lymphocytes, ANAE staining density was more faint than that of non-stimulated cells and the percentages of ANAE positive cells were lower than those of E-rosette positive cells. These results indicated that ANAE activity could be one of T cell markers for porcine mature lymphocytes (spot or granular pattern) of spleen, blood, mesenteric lymph nodes and Peyer's Patch and one of markers for porcine monocytes (diffuse pattern).

Acid α-naphthyl acetate esterase assay in murine lymphocytes

Acid α-naphthyl acetate esterase (ANAE) activity was assayed in cell homogenates and in intact cells by an endpoint colorimetric method, in which sodium dodecyl sulfate was used to stop the reaction. Each method of cell disruption and enzyme solubilization tested here caused a partial loss of the ANAE activity in lymphocyte preparations. The majority of the ANAE activity in lymphocytes was found to be membrane bound. The ANAE activity in thymocytes was over two times lower than that obtained for lymph node and spleen lymphocytes. Macrophages were found to contain about 18 times higher ANAE activity than mature lymphocytes.

Staining of pig lymphocytes subpopulations with acid α-naphthyl acetate esterase

The use of α-naphthyl acetate esterase (ANAE) as a T cell marker in some other species and the broad correlation of incidence of ANAE-positivity and E rosette-formation in the pig suggest that ANAE-staining may be a T-cell marker in the pig. However, by studying the staining of lymphocytes within a variety of rosettes in fixed preparations a similar incidence of pig blood lymphocytes were found to be ANAE + among T cells (E rosettes formed in dextran), B cells antiglobulin rosettes) and Fc-γ receptor-bearing B and T cells (EA rosettes in saline and dextran): complement (C′) receptor-bearing cells showed a higher incidence of staining than other lymphocytes. Analysis of staining morphology suggested that certain morphologies within the B and T lineages may be confined to subpopulations. Thus ANAE positivity is certainly not a marker identifying blood T lymphocytes but could be of some value indicating subpopulations of B and T lymphocytes.

The distribution of N-acetyl-β-glucosaminidase, β-glucuronidase, acid α-naphthyl acetate esterase and α-naphthyl acetate esterase in subpopulations of thymocytes, bone marrow cells and other lymphoid organs in mice

1. 1. Thymocytes, bone marrow lymphocytes, as well as lymphocytes from spleen, lymphoid nodes and peripheral blood were obtained from BALB/c mice. Subpopulations of BALB/c bone marrow T-lymphocyte precursors and immature (small) and mature (large) thymocytes, as established by the percentage of terminal deoxynucleotidyl transferase (TdT) and peanut agglutinin (PNA) positive cells, were obtained by centrifugation on discontinuous density gradients. 2. 2. The activities of N-acetyl-β-glucosaminidase (NAG), β-glucuronidase (BG), acid α-naphthyl acetate esterase (ANAE) and α-naphthyl acetate esterase (NAE) were determined by enzymatic assays of cell extracts of the diverse T-lymphocyte subpopulations, in order to follow their evolution with the maturation of the T-lymphocytes in the thymus. 3. 3. These activities were compared with that determined in lymphocytes from spleen, lymphoid nodes and peripheral blood. 4. 4. The glucidases BG and NAG and the esterases ANAE and NAE present a high decrease in their activities from bone marrow T-lymphocyte progenitors to immature thymocytes. 5. 5. BG, NAG and ANAE activities undergo an about 3-fold increase with the evolution of the thymocytes from small to large cells. Whereas the level of the NAE activity decreases (2-fold) with that evolution of the thymocytes. 6. 6. Lymphocytes from spleen and lymphoid nodes exhibit activities of the glucidases and, specially, the esterases marked by higher than those of thymocyte populations. 7. 7. Peripheral blood lymphocytes also present NAG, ANAE and NAE activities higher than in thymocytes, but their BG activity is lower.

Esterase-positive inflammatory cells in human periapical lesions

The relative distribution of acid α-naphthyl acetate esterase staining cells was studied in inflammatory infiltrates of acute (rapidly progressive) and chronic (slowly progressive) inflammatory periapical lesions, selected on the basis of clinical data. Differential cell counts on sections from the acute (10 granulomas) and chronic (3 granulomas and 7 cysts) inflammatory lesions showed that plasma cells were the predominating α-naphthyl acetate esterase-reactive cells (27%) in acute lesions, whereas T lymphocytes predominated in chronic lesions (25%). Macrophages were more abundant in chronic lesions (13%) compared with acute lesions (6%). Large granular lymphocytes (natural killer cells) could not be identified with certainty among the α-naphthyl acetate esterase-positive cells. The results indicate that there is a relationship between the composition of the inflammatory cell infiltrate and the progression of an inflammatory periapical lesion. The relative distribution of acid α-naphthyl acetate esterase staining cells was studied in inflammatory infiltrates of acute (rapidly progressive) and chronic (slowly progressive) inflammatory periapical lesions, selected on the basis of clinical data. Differential cell counts on sections from the acute (10 granulomas) and chronic (3 granulomas and 7 cysts) inflammatory lesions showed that plasma cells were the predominating α-naphthyl acetate esterase-reactive cells (27%) in acute lesions, whereas T lymphocytes predominated in chronic lesions (25%). Macrophages were more abundant in chronic lesions (13%) compared with acute lesions (6%). Large granular lymphocytes (natural killer cells) could not be identified with certainty among the α-naphthyl acetate esterase-positive cells. The results indicate that there is a relationship between the composition of the inflammatory cell infiltrate and the progression of an inflammatory periapical lesion.

Inhibition of non-specific leukocyte esterase activity. Absence of monocyte esterase activity due to phosphoric and thiophosphoric acid ester intoxication

In vitro evaluation of the effect of five insecticidal phosphoric and 11 thiophosphoric acid esters on different, non-specific human leukocytes esterases indicated that most of the organic phosphor compounds studied inhibited the activity of neutral α-naphthylacetate esterase, α-naphthylbutyryl esterase, and naphthol AS acetate esterase, i.e. the monocyte esterases. The extent of inhibition was dose dependent; the inhibiting dose being identical for the various non-specific esterases. Reactivation with Obidoxim was not successful. Monocyte esterase activity in a human survivor of E 605 intoxication was detectable only after serum acetylcholinesterase had returned to normal levels. The organic phosphor compound studied, however, inhibited neither acid α-naphthylacetate esterase nor naphthol AS-D chloroacetate esterase activity.

Blast Crisis of Chronic Myelogenous Leukemia with Tumor Formation Characterized by T-cell Features—A Case Report

We report a case of chronic myelogenous leukemia (CML) associated with pronounced peripheral lymphadenopathy, with the cells having the philadelphia (Phl) chromosome and T-cell features. A 23-year-old man who was diagnosed as having CML and treated with busulfan was admitted to our hospital because of increasing hepatosplenomegaly and pronounced lymphadenopathy. An axillary lymph node biopsy disclosed that the malignant cells formed rosettes with neuraminidase-treated sheep red blood cells (En) (95.0%) and were positive for Leu 1 (91.8%). Of the cytochemical reactions, peroxidase was negative and periodic acid-Shiff, acid alpha-naphthyl acetate esterase and beta-glucuronidase were all positive. The karyotype of the bone marrow cells was 46 XY Phl positive (22q-), and that of the lymph node cells was 51 XY Phl positive +8, +9, +18, +19, +21, 22q-. He was treated with various anti-leukemic agents and irradiation. Despite such treatments, he died of pneumonia. This is a report of a CML patients with blast crisis and tumor formation characterized by T-cell features.

Inflammatory cells and their subsets in lesions of juvenile periodontitis. A family study.

The inflammatory cells in the gingival biopsy samples from a total of 9 patients with juvenile periodontitis (JP) and from 10 of their family members (JPrelat) belonging to 5 different families were subjected to phenotypic characterization. Plasma cells and their immunoglobulins were stained with immunoperoxidase kits for IgA, IgG, and IgM. B, T, and MPS (cells of the mononuclear phagocyte system) cells were demonstrated by the alpha-naphthyl acetate esterase (ANAE) technique in cryostat sections. The subsets (T helper/inducer and T suppressor/cytotoxic cells) of T cells were detected with monoclonal antibodies OKT4 and OKT8, respectively, using indirect immunofluorescence. Similar studies were completed for 19 age- and sex-matched periodontally healthy subjects. IgG plasma cells for outnumbered the IgA and IgM cells in all three series and were most pronounced in JPrelat series. Most of the inflammatory cells were ANAE-negative (B cells) in all series (78-87%). The highest proportions of T and MPS cells were found in the JP series (10% for both). A statistically significantly higher ratio for TH/Ts (OKT4+/OKT8+) was found in both the JP and JPrelat series as compared with that of the healthy controls. The findings suggest that imbalance (either inherited or acquired) in the immune regulation may play role in the development of JP.

Cellular infiltrate in situ and response kinetics of human intradermal and epicutaneous tuberculin reactions

Monoclonal antibodies used in the avidin-biotin-peroxidase complex (ABC) method and a histochemical azo-dye method for acid α-naphthyl acetate esterase (ANAE) were used to identify T lymphocytes and their functional subpopulations, B cells, and mononuclear phagocytes in tuberculin-test reactions. At 6 and 24 hr up to 52% of all dermal inflammatory cells in situ were T6+, whereas at 72 hr no T6+ cells were observed in the dermis. This suggests that in the initial phases of tuberculin reactions the epidermal Langerhans cells are mobilized to the perivascular spaces in the dermis. At 6, 24, and 48 hr T3+ T lymphocytes usually formed the main inflammatory cell type in situ. The high proportion of Ia+ T lymphocytes and the variations in the local proportion and numbers of T4+ and T8+ cells suggest that T cells play an active role in the generation of a positive tuberculin reaction. At 72 hr T-pattern ANAE+ lymphocytes accounted for 64 ± 9 and 35 ± 10% of all cells in situ in epicutaneous and intradermal tuberculin reactions, respectively ( P < 0.05). The corresponding values for M-pattern ANAE+ macrophages were 27 ± 9 and 53 ± 10% ( P < 0.10). This indicates that the recruitment of mononuclear phagocytes is delayed if tuberculin is applied epicutaneously rather than injected intradermally.