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Biomedical subjects

A Leprini

Publications and source records attributed to A Leprini.

At least 55 records · Page 3Linked to original sources

Identification of autoreactive lymphocytotoxic antibodies in sensitised dialysis and kidney transplant patients.

Autoreactive lymphocytotoxic antibodies, which have been found in sensitised dialysis patients, are generally not considered to be harmful to a renal allograft. In this work the presence of such autoreactive antibodies was investigated in the following groups of sensitised endstage renal disease patients: (a) dialysis patients waiting for a first kidney transplant, (b) kidney transplanted patients, and (c) dialysis patients with a previous failed graft. Only sera from the above patients which showed high reactivity (greater than 30%) against peripheral blood lymphocytes of a random cell panel (R-PBL), were screened for the presence of autoreactive lymphocytotoxic antibodies, by testing at different temperatures against autologous T lymphoblasts (PHA-ATL) and EBV-induced autologous B lymphoblasts (EBV-ABL). The results showed that both blood transfusions, and viral infections such as cytomegalovirus (CMV), correlated with the presence of autoreactive antibodies, and that in addition, by using PHA-ATL and/or EBV-ABL as absorbing reagents, it was possible to remove the antibodies. These absorption procedures allowed the identification of the presence of autoreactive antibodies alone or in combination with other alloantibodies.

Adolescent↗

Large granular lymphocytes have a promoting activity on human peripheral blood erythroid burst-forming units.

Peripheral blood mononuclear cells were fractionated according to the expression of a variety of surface markers, and the fractions obtained were tested for erythroid burst-forming unit (BFU-E) colony formation. BFU-Es were detected in the HLA-DR+ non-T cell fraction, but gave rise to optimum colony numbers only in the presence of a nonadherent, relatively radioresistant cell. This accessory cell was found among the HLA-DR- non-T, non-B cells, a fraction that was particularly enriched in large granular lymphocytes (LGLs). Experiments carried out to assess directly the surface markers of the accessory cell revealed an FcR+, OKM1+, Leu 7+, Leu 11+, OKT4-, OKT8- surface phenotype, which is consistent with that of the majority of LGLs. Peripheral blood LGLs, purified by Percoll density gradient, proved very efficient in promoting optimal BFU-E colony formation. All of these results indicate that LGLs have a potent erythroid burst-promoting activity. Such activity is probably mediated through the release of soluble factors, as shown by the observation that LGL culture supernatants were as effective as LGLs in sustaining colony formation.

Antigens, Surface↗

Granule formation in large granular lymphocytes (LGL). Ultrastructural and cytochemical studies on cells from patients with abnormally expanded lgl populations.

The peripheral blood mononuclear cells from five patients with abnormally expanded populations of large granular lymphocytes (LGL) of possible malignant origin have been studied by electron microscopy. These cells had fewer granules than normal circulating LGL but displayed a variety of organelles possibly involved in the process of granule formation. This process apparently originated from the fusion of vesicles that had a characteristic cup-shape (and appeared to be derived from the Golgi apparatus) with smaller vesicles, mostly of the coated type. This fusion resulted in the formation of multivesicular bodies (MVB) whose limiting membrane was constituted by that of the cup-shaped vesicle. MVB gradually matured into electron-dense granules. A number of cup-shaped vesicles were found positive for the cytochemical localization of thiamine pyrophosphatase (TPP) activity that is a specific marker of the trans aspect of the Golgi apparatus, a finding which confirmed that they were Golgi-derived. The smaller, mostly coated, vesicles were positive for acid phosphatase (AP) and evidence was obtained indicating that they subserved the function of transporting acid hydrolases from the rough endoplasmic reticulum to the cup-shaped vesicles or to the MVB. The plasma membrane did not appear to contribute to MVB or granule formation since endocytic vacuoles induced by reacting the cell surface with the appropriate monoclonal antibodies (followed by peroxidase labelled anti-immunoglobulin) were never seen while fusing with granules or MVB. Rather, these vesicles formed secondary lysosomes after having encountered smooth or, more rarely, coated vesicles. Granulogenesis in LGL, therefore, resembles the process of granule formation observed in secretory cells. The active process of granulogenesis detected in the patient cells and a number of other findings indicate that they were LGL at early stages of maturation. The surface phenotype consistently found in these cells (OKT3+, Leu 1+, OKT8+, Leu 2a+, OKT11+, Leu 5+ and OKM1) is different from that of the majority of the normal LGL and may correspond to that of immature LGL.

Acid Phosphatase↗

Analysis of human peripheral blood lymphocytes isolated by counterflow centrifugation-elutriation.

Human peripheral blood mononuclear cells isolated by Ficoll-Hypaque density gradient centrifugation have been fractionated by counterflow centrifugal elutriation (CCE). Six CCE fractions were obtained and subsequently analyzed as for their content of monocytes, T cells, NK cells and B cells. The various cell types were identified through the expression of specific surface membrane determinants or by cytochemical staining for alpha-naphthyl acid esterase (ANAE). Monocytes were elutriated at the highest counterflow rates whereas the majority of B cells were collected at the lowest counterflow rates. T cells as well as NK cells were mostly concentrated in the intermediate fractions. No differences in the elutriation profile of T cells with the helper-inducer or with the cytotoxic-suppressor surface phenotype were observed. However, the percentages of T cells as determined by surface marker expression decreased with increasing counterflow rates, whereas the percentage of ANAE-positive T cells increased. Yet, T cells recovered at the high counterflow rates had ANAE-reactive organelles larger than those of T cells collected at low counterflow rates. These findings suggest that T cells at different maturational stages could be separated by CCE.

B-Lymphocytes↗

Expression of a receptor for sheep erythrocytes by B lymphocytes from a chronic lymphocytic leukemia patient.

A patient with a B-cell chronic lymphocytic leukemia, whose lymphocytes also formed rosettes with sheep red cells, is described. The B-cell nature of the malignant lymphocytes was determined by surface marker analysis, and cytochemical and ultrastructural studies. The lymphocyte membrane immunoglobulin (IgG1K) did not have anti-sheep red cell activity and was not responsible for the binding of sheep erythrocytes to the leukemic cells as shown by (i) the failure to inhibit rosette formation with anti-immunoglobulin reagents and (ii) the different sensitivity to proteolysis of the membrane immunoglobulin and the sheep erythrocyte receptor. The malignant lymphocytes expressed a receptor for sheep erythrocytes similar to that of normal T cells since they stained with monoclonal antibodies directed against the sheep red cell receptors. Furthermore these antibodies blocked rosette formation. Endogenous labeling experiments demonstrated that the patient's cells produced IgG both of the membrane and of the secretory type. The latter molecular form was also actively secreted. Ultrastructural studies demonstrated that the malignant clone comprised cells at different maturational stages and with different secretory properties. These findings were confirmed by the analysis of intracytoplasmic acid hydrolases, which are normally expressed at late maturational stages. These data are consistent with the hypothesis that a process of maturation was occurring within the malignant clone.

Aged↗

Complement receptor distinguishes between two subsets of large granular lymphocytes with different natural killer activity and cytochemical and ultrastructural features.

Human peripheral blood large granular lymphocytes (LGL)--that is, cells with intracytoplasmic azurophilic (electron-dense) granules, with a positivity for the cytochemical localization of certain acid hydrolases, and with avid surface receptors for the Fc portion of IgG--have been purified on Percoll density gradients. Approximately 30% of these cells expressed receptors for the third complement component (C3R). They were separated into C3R-positive and C3R-negative cells. C3R+ cells had a significantly greater natural killer (NK) activity against K562 target cells than C3R+ cells. This difference was unrelated to the presence in the C3R+ cells of a contaminant cell type incapable of NK activity, since cytochemical and ultrastructural analysis revealed that C3R+ and C3R- fractions contained comparable LGL numbers. Agarose cytotoxicity assays at the single-cell level demonstrated that C3R+ LGL contained a large number of cells that bound to but did not lyse the target. The remaining fully cytotoxic C3R+ LGL had, however, the same killing and recycling properties as the cells from the C3R fraction. Electron microscopy and cytochemical studies showed that C3R+ cells had fewer electron-dense granules than C3R cells and stained more faintly for the localization of alpha-naphtyl acetate esterase. In contrast to C3R cells, C3R+ LGL displayed morphological features suggesting that an active process of granule formation was taking place. Taken together, the data indicate that C3R+ cells represent a discrete subset or a maturational stage of LGL.

Antibodies, Monoclonal↗

PHA-induced human T cell colony formation: enhancing effect of large granular lymphocytes.

The capacity of phytohemagglutinin (PHA)-stimulated human T cells to develop into colonies in agar has been evaluated in the presence or absence of a variety of peripheral blood mononuclear cell subsets. A subpopulation of non-T, non-B cells with receptors for the Fc portion of IgG (i.e. third population cells or TPC) was found to enhance considerably the T cell colony forming capacity. Since TPC have been previously shown to be highly enriched for large granular lymphocytes (LGL, i.e. cells with cytoplasmic azurophilic granules and acid hydrolases), LGL were purified on Percoll density gradients and tested for their T cell colony enhancing capacity. It was shown that LGL were indeed the cells capable of enhancing the T cell colony formation.

Cell Separation↗

Immunofluorescent and ultrastructural analysis of plasma cell degeneration in the chicken Harder's gland.

In this study we describe some aspects of plasma cell degeneration in the chicken Harder's gland. The immunofluorescent patterns of cytoplasmic immunoglobulin (cIg) localization have been studied in relation to the ultrastructure of maturing and degenerating B cells. It appears that Russell body formation through the accumulation of Ig within the cisternae of the rough endoplasmic (RER) does not represent the only mechanism for the formation of cytoplasmic vacuoles. Mitochondrial swelling and disruption or dilation of Golgi cisternae, often preceding alterations of the RER, may be the origin of some vacuoles. It also appears that, in the Harder's gland, degeneration may occur not only in mature plasma cells but also in maturing B cells at a stage when only clusters of polyribosomes are found in the cytoplasm and no RER is yet developed. These observations are relevant to some immunofluorescence and ultrastructural patterns observed in human B-cell pathology.

Animals↗

Receptors for the third complement component on a proportion of large granular lymphocytes from human peripheral blood.

Large granular lymphocytes (LGL) are nonadherent cells with cytoplasmic azurophilic granules, avid receptors for the Fc portion of IgG, and a paranuclear localization of alpha-naphthyl acid esterase or acid phosphatase. LGL constitute the bulk of TG cells (cells with receptors for sheep erythrocytes and for IgG molecules) and null cells (non-T, non-B cells). In the present study we demonstrate that 20-33% of the circulating human LGL express receptors for the third complement component (C3R). When TG cell or null cell fractions from normal individuals or non-T cells from a patient with infantile agammaglobulinaemia (which contained almost exclusively LGL) were rosetted with erythrocytes coated with antibody and complement, a variable number of C3R-bearing cells were detected. Such cells were isolated and analysed further; the great majority of them displayed the cytochemical and ultrastructural features of LGL.

Acid Phosphatase↗

Acid hydrolases as markers of maturation in B-cell chronic lymphocytic leukemia.

Malignant lymphocytes from 30 B-cell chronic lymphocytic leukemia (B-CLL) patients were studied for the cytochemical localization of two acid hydrolases, alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (AT). The large majority of the cells stained for both ANAE and AP in 7 cases, for AP only in 18 cases, and were negative for both the enzymes in 5 cases. Ultrastructural analysis revealed that the cells that displayed more mature morphological features, such as well developed smooth and rough membrane compartments, were those positive for acid hydrolases. That ANAE and AP are expressed by B cells at late stage of maturation was confirmed by the finding that some lymphocytes and all of the plasmacytoid lymphocytes and plasma cells from Walderström's macroglobulinemia, from mixed cryoglobulinemia, and from multiple myeloma patients stained strongly for both ANAE and AP. Using the expression of acid hydrolases and certain ultrastructural features as markers of cell differentiation, it was possible to demonstrate a process of maturation within the single B-CLL clones with accumulation of the cells at stages that differed in the various cases.

Acid Phosphatase↗

Large granular lymphocytes in human peripheral blood: ultrastructural and cytochemical characterization of the granules.

Large granular lymphocytes (LGL) are defined as nonadherent mononuclear cells with cytoplasmic azurophilic granules, avid receptors for the Fc portion of IgG, and cytotoxic functions (NK or ADCC activities). In the present study, the granules of LGL isolated from human peripheral blood have been analyzed by enzyme cytochemistry and electron microscopy. It had been found that: (1) in the single cells, granules at different stages of maturation could be detected: in addition, packaging of the granules took place in the proximity of the Golgi apparatus, which is similar to that seen in secretory cell types. (2) Acid phosphatase (AP) was observed within the granules and the vesicles located in the Golgi area: the Golgi apparatus identified through its thiamine pyrophosphatase-positivity was consistently negative for AP. (3) Alpha naphthyl-acetate esterase (ANAE) activity was localized in the granules as well as on the membrane of LGL and monocytes. (4) The ANAE activity of LGL was of the monocytic and not of the lymphocytic type, as shown by NaF inhibition. (5) The LGL granules, although identifiable as primary lysosomes, were not involved in the process of phagocytosis, since LGL failed consistently to ingest latex particles or opsonized red cells.

Acid Phosphatase↗

Ultrastructural localization of alpha-naphthyl acid esterase in human TM lymphocytes.

The localization of alpha-naphthyl acid esterase (ANAE) activity in T lymphocytes with receptors for IgM (T(M) cells) have been studied at the electron microscope. The electron-opaque product of the cytochemical reaction was detected around, but never inside, single (or groups of) vesicles, which suggested a possible membrane localization of the enzyme activity. These same vesicles were found to contain acid phosphatase by both light- and electron-microscopic examination and were bound by unit membranes; this data indicates that they likely represent primary lysosomes. The presence of such lysosomes in a restricted paranuclear area is a distinctive feature of T(M) cells.

Acid Phosphatase↗

Ultrastructure and cytochemistry of human peripheral blood lymphocytes. Similarities between the cells of the third population and TG lymphocytes.

The ultrastructural and cytochemical features of human peripheral blood TG cells (T cells with receptors IgG) and of the cells of the so-called third population (non-T, non-B cells with high avidity receptors for IgG) have been investigated and compared. Both TG and third-population cells (TPC) contained acid hydrolases with a paranuclear localization of alpha-naphthyl acid esterase, beta-glucuronidase or acid phosphatase. At the electron microscopy level, TG and TPC were indistinguishable and displayed rough cell surface, indented nuclei, abundant cytoplasm with predominance of the smooth over the rough membranes and peroxidase-negative granules. A large proportion of cells of the TPC could form rosettes with sheep erythrocytes after treatment with neuraminidase. The observed close similarities between TG and TPC may suggest that both cell types belong to a special subset of T cells. However, the alternative hypothesis that both TG and TPC are part of a subset unrelated to T cells, such as a new non-T, non-B cell population, or even of the monocytic-macrophage lineage, is also discussed.

Histocytochemistry↗

An experimental study to examine the patency and tissue response of two types of biosynthetic graft used as a replacement for porcine inferior vena cava.

This experimental study has been carried out to evaluate biosynthetic grafts as vascular substitutes. Tubular segments of 35 x 8 mm made of (1) tanned ovine collagen and integral polyester mesh, either of the first (Omniflow I) or second generation (Omniflow II), or (2) polytetrafluoroethylene (e-PTFE), have been sutured in the infrarenal inferior vena cava of pigs, and removed 1 hour, 7, 14, 28, 56 and 112 days after implantation. The patency rate of biosynthetic grafts was higher than that of e-PTFE grafts (p < 0.01). There was no significant difference between the patency of the first generation and second generation collagen grafts. These results indicate that biosynthetic prostheses may be suitable vascular substitutes in low flow and low pressure systems. Improvements in the collagen inner cover structure (Omniflow II vs. Omniflow I), producing greater mechanical endurance, did not enhance long-term patency or the healing patterns of biosynthetic grafts.

Animals↗

[Analysis of the cellular infiltrate and epithelial class I and II molecular expression in edematous type nasal polyps].

Although in the last few years histopathological, immunohistochemical and immunological studies on nasal polyps have been carried out by several Authors, the etiology of these formations still remains unknown. Nasal polyps have a very characteristic structure and have been classified in three histologic types: edematous, glandular and fibrous. In the present report, 11 nasal polyps of edematous type, representing 61% of total number of collected polyps, were studied employing immunohistochemical methods. All the examined polyps were similar in histology and positivity pattern for HLA molecule expression. The edematous core appeared infiltrated (149 cells/mm.2) mainly by eosinophils (> 90%), whereas the peripheral subepithelial connective tissue contained cellular clusters (416 cells/mm2.) made up of different subsets of hematic cells (30.8% were monocytes-macrophages and 48.6% were lymphocytes largely represented by CD4+ cells). On the contrary, mast cells were quite rare (on the average 1.7 cells/mm2.) and located near T cell clusters. The epithelial positivity for HLA-DR and HLA-A,B,C molecules showed a characteristic discontinuous pattern. In the same patient, controlateral nasal mucosa showed a histological structure very similar to that of polyps. The above data suggest that the presence of polyps is the result of an inflammatory process brought about by a complex array of cellular and humoral components.

Edema↗