Prothymosin α receptors on peripheral blood mononuclear cells

Cordero, Oscar J.; Sarandeses, Concepción S.; Nogueira, Marco Aurélio

doi:10.1016/0014-5793(94)80233-5

Cited by 30 publications

(18 citation statements)

References 24 publications

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“…Rabbit anti‐(α‐ l ‐fucosidase) Ig was radiolabeled with Na 125 I (DuPont NEN, Germany, or Amersham PharmaciaBiotech) by using Iodobeads (Pierce, Rockford, IL), a modification of the Chloramine‐T method, following the method previously described [24]. Fresh or cultured cells were resuspended in isotonic NaCl/P i and stained with 125 I‐labeled rabbit anti‐(α‐ l ‐fucosidase) Ig for 30 min as described above.…”

Section: Methodsmentioning

confidence: 99%

“…The solubilized membrane proteins, lyophilized if necessary for protein concentration, as well as whole‐cell lysate samples or commercial α‐ l ‐fucosidase were diluted in the electrophoresis sample buffer and analyzed on an SDS/PAGE (10% in the first case, 12% in the others) as described [21,24] before processing for immunoblot assay on Hybond‐P poly(vinylidene difluoride) membranes (Amersham PharmaciaBiotech, Rainham, UK) by using rabbit anti‐(α‐ l ‐fucosidase) Ig, 1 : 100 (v/v) from the antisera Ig fraction (1 : 500 in the Ig characterization experiments). The specific Ig was revealed with goat anti‐(rabbit Ig) Ig labeled with horseradish peroxidase and ECL (Amersham PharmaciaBiotech), or with goat anti‐(rabbit Ig) Ig coupled to alkaline phosphatase (1 : 1000 dilution) and 5‐bromo‐4‐chloro‐3‐indolyl phosphate (BCIP) and Nitro Blue Tetrazolium (NBT) substrates.…”

Section: Methodsmentioning

confidence: 99%

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Cell surface human α‐L‐fucosidase

Cordero

Merino

Cadena

et al. 2001

European Journal of Biochemistry

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The acid a-l-fucosidase is usually found as a soluble component of lysosomes where fucoglycoconjugates are degraded. In the present investigation, we have demonstrated the existence of a cell surface protein with enzymatic a-l-fucosidase activity that crossreacts specifically with a rabbit anti-(a-l-fucosidase) Ig. By different approaches, this a-l-fucosidase, which represents 10±20% of the total cellular fucosidase activity, was detected in all the tested human cells (hemopoietic, epithelial, mesenchymal). Two bands of < 43±49 kDa were observed, although theoretical data support the possibility of having the same genetic origin that the known 50 to 55-kDa M r a-l-fucosidase. We speculate about an alternative traffic pathway for the plasma membrane a-l-fucosidase to work on the rapid turnover of glycoproteins.Keywords: a-l-fucosidase; plasma membrane; protein traffic; CD26; glycoprotein turnover.The lysosomal a-l-fucosidase (EC 3.2.1.51) is involved in the hydrolytic degradation of fucose-containing molecules. Mammalian a-l-fucosidases are multimeric forms of glycoproteins of < 53 kDa exhibiting optimal activity between pH values of 4 and 6.5 [1]. There appears to be a considerable degree of structural heterogeneity, both tissue-specific and within the same tissue. This is probably due to a different content in sialic acid residues [2,3], in addition to the two different alleles and genetic polymorphism encoded by the FUCA1 gene [4,5]. The significance of the FUCA2 gene is currently unknown [4±6].The absence or gross deficiency of a-l-fucosidase activity causes accumulation of fucoglycoconjugates, which leads to the genetic neurovisceral storage disease fucosidosis in mammals [7,8]. A new syndrome, previously known as leukocyte adhesion deficiency II, has been recently characterized as a generalized metabolic disease consisting of severe hypofucosylation of glycoconjugates [9]. Moreover, an abnormal intracellular and extracellular distribution of a-l-fucosidase is found, for example, in cystic fibrosis (decreased levels in antiserum and higher levels in skin fibroblasts) or in colon carcinomas (decreased levels in antiserum and in tumor tissue) [10±13]. In general, tissue differentiation and development through cell±cell recognition are modulated by sequential changes of the sugar chains of cell surface glycoproteins. As the expression or deletion of a-fucosyl residues linked at various positions of sugar chains of glycoproteins is one of these alterations, the role of a-l-fucosidase in these processes is of considerable interest.The majority (90±100%) of the a-l-fucosidase activity in almost all mammalian tissues investigated is in the soluble fraction [1], the human brain being the exception [14]. A serum form of a-l-fucosidase, which has attracted interest as a diagnostic marker in many clinical studies [15], has turned out to be similar to the tissue forms [4±6,16,17]. Very recently, a-l-fucosidase from rat testis and epididymal spermatozoa was found by immunocytochemistry to be associated with the outer p...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Cell surface human α‐L‐fucosidase

Cordero

Merino

Cadena

et al. 2001

European Journal of Biochemistry

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“…Receptors for Pro ~1 were found on lymphocytes and NK-like cell lines (Cordero et al 1994). Pro cd mainly affects the IL-2/IL-2 receptor system and this may contribute to the stimulation of cytotoxicity against cultured tumor cells (Baxevanis et al 1993).…”

Section: Discussionmentioning

confidence: 99%

“…Pro ~.1, a fully defined protein of 12 kDa, demonstrated in vitro immunostimulating effects on different cell types, including NK cells, T lymphocytes, granulocytes and monocytes (Cordero et al 1990(Cordero et al , 1992Wykretowicz et al 1994;Garbin et al 1994a;Gast et al 1995). Binding sites for Pro c~l were found on human peripheral mononuclear cells, including NK-like lymphocytes (Cordero et al 1994). Pro ctl increased NK cytotoxicity against different tumor cells, promoted the production of IL-2 and IFN7 and increased the expression of the membrane receptor for IL-2.…”

Section: Introductionmentioning

confidence: 96%

Interleukin-2-activated killer cell activity in colorectal tumor patients: evaluation of in vitro effects by prothymosin?1

Eckert

Grünberg

Immenschuh

et al. 1997

J Cancer Res Clin Oncol

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The effects of prothymosin alpha1 (Pro alpha1) in combination with interleukin-2 (IL-2) on peripheral blood lymphocytes from 50 colorectal tumor patients at different stages were studied with respect to immunocytotoxicity, adhesion to cultured SW620 colon carcinoma cells, secretion of cytokines and expression of adhesion and surface marker molecules. On average, the patients showed lower natural killer (NK) cell activity than healthy donors, which was associated with a lower adhesion capacity to the tumor target cells. The NK cell activity of the patients was inversely related to the tumor stage. The generation of lymphokine(IL-2)-activated killer (LAK) cell activity was found to be comparable on lymphocytes from healthy individuals and patients and was not correlated to tumor stage. Pro alpha1 stimulated patients' LAK cell activity only, primarily at the early stage (Dukes A/B). The Pro alpha1 effect was associated with an increased adhesion of lymphocytes to tumor target cells and an increased secretion of the deficient IL-2-induced IFN gamma secretion. No significant effects on the low level of TNF alpha secretion was noted. By flow cytometry, Pro alpha1 in combination with IL-2 augmented the expression of the NK cell markers CD56, CD16/56, the subset CD3/16/56 and CD25 on lymphocytes of the patients. In contrast, Pro alpha1 was equally effective by increasing the expression of CD18 and CD11a on lymphocytes from the patients and from normal controls. In conclusion, Pro alpha1, in combination with IL-2, can partially normalize lymphocyte deficiencies of colon cancer patients in vitro. This potential might provide an experimental basis for applying Pro alpha1 or related thymic peptides in selected immunotherapies against colorectal tumors.

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“…Buffy coats from healthy donors were kindly provided by the Centro de Transfusiones de Galicia, Santiago, or Banco de Sangre, Hospital General Vall d'Hebrón, Barcelona, Spain. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll Paque PLUS (Pharmacia, Uppsala, Sweden) density gradient centrifugation [21] and cultured at 10 6 cells/mL in RPMI 1640 (Sigma) supplemented with 10% inactivated fetal calf serum (FCS; Gibco BRL, Grand Island, NY), 100 g/mL of streptomycin, and 100 IU/mL of penicillin (Sigma) in a 5% CO 2 humidified atmosphere. PBMCs were activated with 1 g/mL of PHA-P or with 0.5 g/mL of anti-CD3 in the presence or absence of cytokines.…”

Section: Cell Isolation and Culturementioning

confidence: 99%

Cytokines regulate membrane adenosine deaminase on human activated lymphocytes

Cordero

Salgado

Fernández-Alonso

et al. 2001

Journal of Leukocyte Biology

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CD26 is a lymphocyte marker that can anchor adenosine deaminase (ADA) on the T cell surface. We found that ADA is regulated by cytokines on the cell surface during T cell activation. By means of flow cytometry, immunofluorescence, and immunoblotting techniques, we found that interleukin (IL)-2 and IL-12 up-regulate ecto-ADA and CD26 expression. In clear contrast, IL-4 led to down-regulation of lymphocyte surface ADA without modifying the level of CD26. Moreover, neither circulating ADA transcription nor mRNA translation was regulated by cytokines. These results, along with absence of total-ADA modulation, the variable amount of ADA found in purified plasma membranes, and the different effect of Brefeldin A on the surface presence of ADA and CD26 indicated that cytokines regulate the translocation of ADA towards the cell surface through a mechanism not involving CD26. Ecto-ADA protected activated lymphocytes from the toxic effects of extracellular adenosine. Therefore, this cell surface ADA control might constitute part of the fine immunoregulatory mechanism of adenosine-mediated signaling through purinergic receptors in leukocytes.

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Prothymosin α receptors on peripheral blood mononuclear cells

Cited by 30 publications

References 24 publications

Cell surface human α‐L‐fucosidase

Cell surface human α‐L‐fucosidase

Interleukin-2-activated killer cell activity in colorectal tumor patients: evaluation of in vitro effects by prothymosin?1

Cytokines regulate membrane adenosine deaminase on human activated lymphocytes

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