Positive Cell Types Research Articles

Characterization of NADPH-diaphorase-positive glial cells of the tench optic nerve after axotomy.

NADPH-diaphorase (ND) positive cell types were characterized throughout the optic nerve of the tench in normal conditions and after optic nerve transection with survival periods of 1, 3, 7, 14, 30, 60, 120 and 180 days. Astrocytic markers (S100 and glutamine synthetase) and the microglial marker tomato lectin were employed. In the control prechiasmatic optic nerve two types (types I and II) of ND-positive glial cells appeared. All type I cells showed S100 immunoreactivity, whereas only a subpopulation of them were positive to glutamine synthetase. Type II cells only presented S100 immunoreactivity. In the control anterior optic tract, all ND-positive glial cells (type III) presented immunolabeling to S100 and glutamine synthetase. After transection, types I and II did not show any changes in the staining patterns for the glial markers when observed. Two new types of ND-positive glial cells (types IV and V) were observed after axotomy. All type IV cells were S100-immunopositive, and a subpopulation presented glutamine synthetase immunolabeling. Only a subpopulation of type V cells showed glutamine synthetase immunostaining. The presence of type IV or V cells in the lesioned optic nerve occurred simultaneously with significant decreases or absence of type I cells. These data suggest that type I and III cells are astrocytes and type II cells are oligodendrocytes. Types IV and V cells are the result of the activation of type I cells after optic nerve section. The polymorphism observed in ND-positive cells may reflect different cell functions during degenerative and regenerative processes.

LIF Upregulates Expression of NK‐1R in NHBE Cells

Leukemia inhibitory factor (LIF), a cytokine at the interface between neurobiology and immunology, is mainly mediated through JAK/STAT pathway and MAPK/ERK pathway. Evidence suggested LIF is related to the higher expression of neurokinin-1 receptor (NK-1R) in asthma. In this study, the immunohistochemistry stain showed the expressions of NK-1R, LIF, p-STAT3, and p-ERK1/2 in the lung tissues of allergic rats were increased compared with the controls, and the main positive cell type was airway epithelial cell. Normal human bronchial epithelial cells were treated with LIF in the presence or absence of AG490 (JAK2 inhibitor), PD98059 (MEK inhibitor), and the siRNA against STAT3. Western blot and RT-PCR indicated that LIF induced the expression of NK-1R, which was inhibited by the inhibitors mentioned above. No significant interaction was found between JAK/STAT pathway and MAPK/ERK pathway. In summary, bronchial epithelial cell changes in asthma are induced by LIF which promotes the expression of NK-1R, and JAK/STAT pathway and MAPK/ERK pathway may participate in this process.

Neuropilin-1 Expression by Endothelial Cells and Retinal Pigment Epithelial Cells in Choroidal Neovascular Membranes

To determine if vascular endothelial growth factor (VEGF) coreceptor neuropilin-1 (NP-1) is expressed in choroidal neovascularization (CNV) and to localize the expression. Laboratory investigation. Six CNV membranes (CNVMs) obtained from patients with subfoveal CNV attributable to age-related macular degeneration (AMD) underwent immunohistochemistry for VEGF receptor-2 (VEGFR-2) and NP-1. The positive cell types were identified by double staining with anticytokeratin, anti-CD31, and antismooth muscle actin (SMA). Immunohistochemical staining revealed positivity for VEGFR-2 and NP-1 in all six CNVMs. Both receptors were strongly expressed by new choroidal endothelial cell forming vessels and CD-31-positive cells in the nonvascular area. They were also expressed in the pigmented retinal pigment epithelial layer and by cytokeratin-positive, nonpigmented retinal pigment epithelium cells in the nonvascular area. The nonpigmented retinal pigment epithelium cells positive for VEGFR-2 and NP-1 were highly colocalized with SMA. The presence of both VEGF and NP-1 suggest that NP-1 may play a role in the evolution of CNV in AMD.

Human Herpesvirus 8 Infection in Patients With Cutaneous Lymphoproliferative Diseases

To investigate the prevalence of human herpesvirus 8 (HHV-8; Kaposi sarcoma-associated herpesvirus) infection in patients with lymphoproliferative skin diseases such as large-plaque parapsoriasis (LPP) and mycosis fungoides compared with inflammatory cutaneous conditions or healthy control subjects. A survey study was undertaken in 123 subjects with various clinical conditions. All patients had been seen in the Dermatology Department of the San Gallicano Dermatology Institute, Rome, Italy, in the last 2 years. Forty-five patients with inflammatory or autoimmune cutaneous diseases, 50 healthy control subjects, 10 patients with LPP, 12 patients with mycosis fungoides, and 6 patients with classic Kaposi sarcoma were included in the study. The prevalence of HHV-8 infection was investigated with serologic studies using the gold standard assay based on body cavity-based B-cell lymphoma-1 cells latently infected with HHV-8. The presence of HHV-8 conserved sequence, corresponding to open reading frame 26, was also assessed in the peripheral blood and lesion tissue samples from patients with lymphoproliferative cutaneous diseases with nested polymerase chain reaction. The presence and distribution of cell types infected with HHV-8 in the lesion tissues was determined with immunohistochemical staining with the monoclonal antibody directed against the latent nuclear antigen-1 of HHV-8 encoded by open reading frame 73. In healthy control subjects and patients with inflammatory skin diseases, 13.9% were found to have antibody against HHV-8, consistent with the seroprevalence population in Italy. A highly significant association of HHV-8 infection and LPP was found (100%) compared with mycosis fungoides (25%). The peripheral blood mononuclear cells in 8 of 10 patients with LPP were found to harbor viral sequences at nested polymerase chain reaction, whereas none of them had a detectable serum viral load. All LPP lesion tissue samples were positive for HHV-8-encoded open reading frame 26, and the presence of HHV-8-infected cells was confirmed by immunohistochemistry profiles performed on paraffin-embedded tissues from 4 of 10 patients. The positive cell types included endothelial cells and the infiltrating dermal lymphocytes, characteristic hallmarks of LPP. Analysis of T-cell receptor gamma chain rearrangements in lesion tissue from our patients confirmed the lack of a significant association between T-cell clonality and LPP. These data suggest that HHV-8 may play a role in the onset of LPP, a disease whose cause and evolution are still undefined and which has often been considered the early stage of mycosis fungoides.

Lymphoid Differentiation Pathways Can Be Traced by TCR δ Rearrangements

TCR gene rearrangement generates diversity of T lymphocytes by V(D)J recombination. Ig genes are rearranged in B cells using the same enzyme machinery. TCRD (TCR delta) genes are frequently incompletely rearranged in B precursor leukemias and recently were found in a significant portion of physiological B lymphocytes. Incomplete TCRD rearrangements (V-D) thus serve as natural indicators of previous V(D)J recombinase activity. Functional V(D)J recombinase has recently been found in murine NK precursors. We tested whether physiological NK cells and other leukocyte subpopulations contained TCR rearrangements in humans. This would provide evidence that V(D)J recombinase was active in the ancestry cells and suggest common pathways among the positive cell types. TCRD were rearranged in 3.2-36% of NK cells but not in nonlymphoid leukocytes. The previously known phenomenon of TCRD transcription in NK cells is a possible mechanism that maintains the chromatin open at the TCRD locus. In comparison, TCRG rearrangements were frequent in T cells, low to negative in B and NK cells, and negative in nonlymphoid cells, suggesting a tighter control of TCRG. Levels of TCRD rearrangements were similar among the B lymphocyte subsets (B1-B2, naive-memory). In conclusion, human NK cells pass through a differentiation step with active V(D)J recombinase similar to T and B lymphocytes and unlike nonlymphoid leukocytes. This contradicts recent challenges to the concept of separate lymphoid and myeloid differentiation.

Targeting Sindbis virus-based vectors to Fc receptor-positive cell types

Some viruses display enhanced infection for Fc receptor (FcR)-positive cell types when complexed with virus-specific immunoglobulin (Ig). This process has been termed antibody-dependent enhancement of viral infection (ADE). We reasoned that the mechanism of ADE could be exploited and adapted to target alphavirus-based vectors to FcR-positive cell types. Towards this goal, recombinant Sindbis viruses were constructed that express 1 to 4 immunoglobulin-binding domains of protein L (PpL) as N-terminal extensions of the E2 glycoprotein. PpL is a bacterial protein that binds the variable region of antibody kappa light chains from a range of mammalian species. The recombinant viruses incorporated PpL/E2 fusion proteins into the virion structure and recapitulated the species-specific Ig-binding phenotypes of native PpL. Virions reacted with non-immune serum or purified IgG displayed enhanced binding and ADE for several species-matched FcR-positive murine and human cell lines. ADE required virus expression of a functional PpL Ig-binding domain, and appeared to be FcγR-mediated. Specifically, ADE did not occur with FcγR-negative cells, did not require active complement proteins, and did not occur on FcγR-positive murine cell lines when virions were bound by murine IgG-derived F(ab′) 2 fragments.

Lithium selectively increases neuronal differentiation of hippocampal neural progenitor cells both in vitro and in vivo.

Lithium has been demonstrated to increase neurogenesis in the dentate gyrus of rodent hippocampus. The present study was undertaken to investigate the effects of lithium on the proliferation and differentiation of rat neural progenitor cells in hippocampus both in vitro and in vivo. Lithium chloride (1-3 mM) produced a significant increase in the number of bromodeoxyuridine (BrdU)-positive cells in high-density cultures, but did not increase clonal size in low-density cultures. Lithium chloride at 1 mM (within the therapeutic range) also increased the number of cells double-labeled with BrdU antibody and TuJ1 (a class III beta-tubulin antibody) in high-density cultures and the number of TuJ1-positive cells in a clone of low-density cultures, whereas it decreased the number of glial fibrillary acidic protein-positive cells in both cultures. These results suggest that lithium selectively increased differentiation of neuronal progenitors. These actions of lithium appeared to enhance a neuronal subtype, calbindin(D28k)-positive cells, and involved a phosphorylated extracellular signal-regulated kinase and phosphorylated cyclic AMP response element-binding protein-dependent pathway both in vitro and in vivo. These findings suggest that lithium in therapeutic amounts may elicit its beneficial effects via facilitation of neural progenitor differentiation toward a calbindin(D28k)-positive neuronal cell type.

Immune Reconstitution and Tolerance after Allogeneic Hematopoietic Stem Cell Transplantation

We have evaluated recovery of CD56 positive and other cell types following allogeneic stem cell transplantation and have found that the recovery of CD56 positive cells was faster than other lymphoid cells after allogeneic stem cell transplantation, while the recovery of CD4 positive cells was markedly delayed. Chimerism analysis showed that mixed chimerism was often observed in younger (<30 years old) patients. Mixed chimerism in older (≥30 years old) patients was associated with rejection and relapse, while this was not found in younger patients. Among the chimerism of various cell populations, donor-derived CD56-positive cells are important in early engraftment when determined in allogeneic nonmyeloablative stem cell transplantation (allo-NST), regardless of the proportion of donor-derived CD3-positive cells. Complementarity-determining region three (CDR3) size spectratyping in T-cell receptor (TCR) chain subfamilies (Vβ) showed that high level of diversity in TCR Vβ repertoire is important for a late rejection and skewed TCR Vβ repertoire is correlated with the occurrence of graft-versus-host disease (GVHD) especially chronic GVHD. Expression of inhibitory natural killer (NK) cell receptors such as CD158b and CD94/NKG2A on peripheral CD3-negative and -positive cells were increased in parallel with GVHD. Interestingly, these cells appeared to control GVHD, while preserving graft-versus-leukemia (GVL) effect. Analysis of cytokine gene expression in peripheral blood mononuclear cells showed that type 1 helper T cells (Th1)-derived cytokines increased in severe GVHD, while Th2-derived cytokines such as IL-4, IL-10 and IL-13 increased in mild GVHD. These results indicate that Th2 cells suppress GVHD, although Th1 cells augment GVHD. Taken together, evaluation of immune reconstitution and tolerance in patients receiving allogeneic stem cell transplantation from the various viewpoints is essential and useful to obtain better clinical outcome.

Comparison of the effectiveness of biphase and monophase rectangular pulses for the inactivation of micro-organisms using pulsed electric fields

The use of pulsed electric fields (PEF) for the inactivation of micro-organisms is currently of considerable interest as an alternative method to the conventional thermal methods of food pasteurization presently in widespread use. Many parameters of PEF influence the effectiveness of inactivation, one of which is the pulse profile, which is the subject of the present study. In this study, a pulse-forming network was designed and constructed to allow direct comparison of the effectiveness of a rectangular pulse (single polarity) with a rectangular pulse (incorporating a rapid reversal of polarity for the second half of the pulse). The pulses of 30 kV/cm in electric field intensity, 2 /spl mu/s in duration for monophase, and 1 /spl mu/s for each phase for biphase, were applied to parallel-plate test cells. Three different species of bacteria were tested including Gram negative and positive cell types. The bacteria were suspended in peptone water solutions of varied peptone concentration and hence, conductivity. The results show that, contrary to previous findings, biphase pulsing did not provide superior inactivation levels compared with monophase pulsing, for all three bacteria tested. Increasing the conductivity of the suspension fluid produced a general trend toward similar levels of inactivation of biphase compared with monophase PEF treatment, but in no case was it found that biphase PEF treatment was superior to monophase PEF treatment.

Biochemical and Morphological Diversity among Folliculo-Stellate Cells of the Mink (Mustela vison) Anterior Pituitary

The folliculo-stellate (FS) cells are agranular cells of the anterior pituitary whose origin and function are still a matter of debate. This study examined the presence, topography, and morphological characteristics of FS cells in the mink anterior pituitary throughout the annual reproductive cycle. The S-100 protein was used as a FS cell marker. Immunoperoxidase labeling on tissue sections demonstrated the presence of two types of S-100 positive cells. Type 1 cells were stellate-shaped cells whose nuclei were localized near the center of pituitary follicles. In this type, S-100 labeling was strong in anterior pituitary sections obtained during spring, a period characterized by high prolactin pituitary content and low gonadotropin pituitary content. Type 2 cells were rounded cells occupying the periphery of the follicles. During periods of low prolactin pituitary content and high gonadotropin anterior content the type 2 S-100 positive cells formed aggregates of several cells. The total number of S-100 positive cells was constant during these two periods of the annual reproductive cycle, suggesting that type 1 and type 2 may reflect different morphological and physiological states of the same cell. Of the two subunits, α and β, that, combined, form three different dimeric S-100 proteins, mink FS cells expressed mostly the β subunit. FS cells also expressed the glial fibrillary acidic protein (GFAP). In culture, 8 ± 3% of anterior pituitary cells were S-100 positive. Cultured S-100 cells were elongated, polygonal, or rounded. The S-100 labeling accumulated in the cytoplasm around and within the nucleus, whereas it was weak in pseudopods and large cytoplasmic vacuoles. The presence of pseudopods suggests that cultured FS cells could migrate. The vacuoles may be related to the phagocytic activity ascribed to these cells. Some FS cells presented membrane blebbing and peripheral vesicles that were immunopositive for S-100 and that may indicate a secretory activity. Cultured FS cells possessed actin filaments organized as a peripheral network; a few actin cables were also observed running across the cytoplasm. Pseudopods depicted a highly organized actin network. The microtubules of FS cells expanded throughout the cytoplasm. The intermediate filaments expressed by cultured FS cells were GFAP and vimentin. GFAP labeling was punctate and vimentin was organized as filaments. All cultured S-100 cells were positive for vimentin, suggesting a mesenchymal origin for the cells, and all cultured S-100 positive cells were positive for GFAP, suggesting a neuroectodermal origin. In conclusion, S-100 positive cells are heterogeneous with respect to cell shape and expression of S-100 subunits in the mink anterior pituitary. The presence of morphologically different S-100 positive cells is modified in accordance with the endocrine status of the animal, suggesting that FS cells may be involved in the modulation of the anterior pituitary endocrine activity in the mink.

Cellular localization of estrogen receptor beta messenger ribonucleic acid in cynomolgus monkey reproductive organs.

There is now evidence that the recently identified estrogen receptor (ER) beta is more widely distributed in the body than is ER-alpha. In order to gain more information about the role of ER-beta in reproduction, we have investigated by in situ hybridization the localization of mRNA expression of this ER subtype in adult monkey reproductive organs. In the pituitary gland of animals of both sexes, in both the anterior and intermediate lobes, a large number of cells were positive. No specific signal was observed in the posterior lobe. In the ovary, granulosa cells in primary and growing follicles highly expressed ER-beta mRNA. The theca interna cells were also strongly labeled. In some corpora lutea, the luteal cells were strongly labeled, while in other ones, the signal was weak. A hybridization signal was also detected in the ovarian surface epithelium. In the uterus, ER-beta mRNA was found in high concentration in glandular epithelial cells and stromal cells of the endometrium, while weaker labeling was consistently observed in smooth muscle cells. In the mammary gland, labeling was detected in the epithelial cells of acini and interlobular ducts as well as stromal cells. In the testis, specific labeling was detected in the seminiferous epithelium whereas the interstitial Leydig cells were unlabeled. Although it was not possible to clearly identify all the positive cell types, it appears that Sertoli cells as well as the vast majority of germinal cells express ER-beta mRNA. In the prostate, the secretory epithelial cells exhibited a specific autoradiographic reaction while the stromal cells did not show mRNA expression. The epithelial cells of the prostatic urethra showed a strong labeling. No hybridization signal was detected in the seminal vesicles. It then appears quite clear that ER-beta is expressed in a cell-specific manner in all the monkey reproductive organs studied. In the female, the wide distribution of these receptors in the ovary and uterus suggests that ER-beta may play an important role in the mediation of the known effects of estrogen in reproduction functions. In the male testis and prostate, ER-beta has been found in cells that contain very little or no ER-alpha. The role of circulating or locally produced estrogens in the male reproductive system remains to be clarified.

Effect of all- trans retinoic acid on chemotherapy induced apoptosis and down-regulation of Bc1-2 in human myeloid leukaemia CD34 positive cells

Acute myeloid leukaemia (AML) is a heterogeneous malignant disease in which disease progression at the level of CD34 positive cells has a major impact in drug resistance and relapse. The multi-drug resistance (MDR1) gene product, P-glycoprotein is expressed mainly in CD34 positive AML cells and Bc1-2 is expressed simultaneously with several putative drug resistance parameters in these cells. Bc1-2 over-expression is associated with CD34 positivity, poor response to chemotherapy and reduced overall survival in AML patients. Recently, all- trans retinoic acid (RA) has been reported to enhance cytarabine-induced apoptosis and downregulate Bc1-2 in several human myeloid leukaemia CD34 negative cells. The two CD34 positive human myeloid leukaemia cell lines: KG1 and KGla have the unique feature of expressing significant functional P-glycoprotein. Thus, the efficacy of RA in enhancing cytrabine- and fludarabine-induced apoptosis and overcoming the resistance was examined in both KG1 (CD34+CD7−) and KGla (CD34+CD7+) human myeloid leukaemia cells in the present study. Both cytarabine and fludarabine induced a dose–dependent increase in the number of apoptotic cells in both CD34 positive cell types. Interestingly, the cytarabine-induced apoptosis was significantly more than fludarabine-induced apoptosis in both cell types. All- trans RA alone failed to induce apoptosis or inhibit proliferation of either of the two human CD34 positive leukaemia cell types. However, RA enhanced cytarabine- or fludarabine-induced apoptosis and inhibition of proliferation in KG1 CD34+CD7− but not in KGla CD34+CD7+ myeloid leukaemia cells. As single agents, RA, cytarabine and fludarabine reduced Bc1-2 expression in a dose–dependent manner in both cell types. Using a quantitative ELISA assay, the Bc1-2 protein concentration was reduced by 86 or 100% after 72 h of treatment with 10 μM cytarabine or fludarabine, respectively, in both CD34 positive leukaemia cell types. The addition of RA to cytarabine enhanced its induced reduction of Bc1-2 in KG1 CD34+CD7− but not in KGla CD34+CD7+ human myeloid leukaemia cells. Meanwhile, RA failed to augment fludarabine-induced reduction of Bc1-2 in both cell types. In conclusion, the present results suggest a potential role for the combination of RA and cytarabine in the treatment of refractory and/or relapsed AML patients with CD34+CD7− but not CD34+CD7+ blast cells.

Primary cell culture of LHRH neurones from embryonic olfactory placode in the sheep (Ovis aries).

The aim of this study was to establish an in vitro model of ovine luteinizing hormone-releasing hormone (LHRH) neurones. Olfactory placodes from 26 day-old sheep embryos (E26) were used for explant culture. Cultures were maintained successfully up to 35 days, but were usually used at 17 days for immunocytochemistry. LHRH and neuronal markers such as neurofilament (NF) were detected by immunocytochemistry within and/or outside the explant. Three main types of LHRH positive cells are described: (1) neuroblastic LHRH and NF immunoreactive cells with round cell body and very short neurites found mainly within the explant, (2) migrating LHRH bipolar neurones with an fusiform cell body, found outside the explant, (3) network LHRH neuron, bipolar or multipolar with long neurites connecting other LHRH neurons. Cell morphology was very similar to that which has been described in the adult sheep brain. These results strongly suggest that LHRH neurones in the sheep originate from the olfactory placode. This mode may represent a useful tool to study LHRH neurones directly in the sheep.

The HNK-1 Carbohydrate Epitope and the Human Eye in Health and Disease.

The HNK-1 carbohydrate epitope is part of many cell membrane and extracellular matrix molecules, several of which have been implicated in cell adhesion. It is a versatile tool in eye research. In the human eye this epitope is present in the retina, the optic and ciliary nerves, the ciliary and iris epithelia, the zonular lamella, and the sclera. It is phylogenetically conserved, but the positive cell types vary from species to species. In addition to revealing interspecies differences in the vertebrate retina, the HNK-1 epitope has been used to identify a novel cell type in the eye: the subepithelial matrix cells that reside in the inner connective tissue layer (ICTL) of the ciliary body. Although these cells resemble fibroblasts in ultrastructure, they form a distinct cell population that differs in antigenic profile from fibroblasts in other tissues. The HNK-1 epitope is also associated with the elastic fiber system of the ICTL, which may be produced by the subepithelial matrix cells. It may help to structurally stabilize the ciliary body and the retina. The HNK-1 epitope is also involved in many important eye diseases. The subepithelial matrix cells seem to be susceptible to irrreversible atrophy as a result of glaucoma, thermal injury, and tissue compression. On the other hand, the HNK-1 epitope is found in the extracellular matrix of secondary cataracts and may contribute to its pathogenesis. Finally, this epitope has proved to be useful in identifying deposits of exfoliation material, and in tracing neuroepithelial derivatives in developmental anomalies and tumors of the eye.

Effect of penclomedine (NSC-338720) on telomere fusions, chromatin blebbing, and cell viability with and without telomerase activity and abrogated p53 function

Telomeres, or chromosome ends, are essential in maintaining chromosomal integrity. Telomeres consist of a short hexameric sequence, 3′-TTAGGG-5′, repeated in tandem arrays added to chromosomes by the ribonucleoprotein enzyme telomerase. In this study, we assessed whether penclomedine, a novel synthetic pyridine compound presently being evaluated in clinical trials for its anticancer activity, influences telomere fusions (chromosome end-to-end associations) and telomerase activity in cells in culture. We found that penclomedine reduced the mitotic index, induced chromosome end associations in all phases of the cell cycle, and rapidly induced chromatin blebbing in a concentration-dependent manner in both cervical carcinoma (HeLa) cells and in normal human fibroblasts (AG1522). However, the effectiveness of the drug was much more pronounced in HeLa cells. In addition, there was a drug-mediated, concentration-dependent decline in telomerase activity noted in the HeLa cells that correlated with a decrease in mitotic index and an increase in telomere fusions. Interestingly, when the mitotic index, chromatin blebbing, and telomere fusions were compared between the telomerase positive (HeLa) and negative (AG1522) cell types, penclomedine affected chromatin stability to a greater extent in those cells with detectable telomerase activity. In addition, telomerase positive colorectal carcinoma cells with abrogated p53 (RC-10.3 cells) were more sensitive to penclomedine than were telomerase positive cells with wild-type p53 (RKO cells). These studies suggest that penclomedine may have a therapeutic advantage in killing tumor cells that are positive for telomerase activity and defective in p53 function.

The role of angiogenic factor bFGF in the development of dural AVFs.

Dural arteriovenous fistulas (dAVFs) are known to be acquired disorders. Angiogenic stimulants, such as basic fibroblast growth factor (bFGF), may be involved in the evolution of this disorder. We examined the appearance and localization of bFGF, in human dAVF sinuses, versus a control using immunohistochemical techniques, to evaluate these qualitative differences. Four human dural sinuses from dAVF patients, and one control dural sinus were removed at surgery or autopsy and used for this study. Immunohistochemistry for bFGF was performed in all five specimens to identify its appearance and localization. Immunohistochemistry for alpha smooth muscle actin, factor VII related antigen, and macrophage (CD-68) were done in all tissues to identify the bFGF positive cell types. In the control dural sinus, there was negative staining by bFGF immunohistochemistry. However, in all four sinuses of the dAVF patients, smooth muscle cells, endothelial cells, and meningeal cells were stained positively in various degrees by bFGF immunohistochemistry. bFGF may be significant in the development of dAVFs, judging from its strong immunoreactivity in the sinuses of dAVF patients.

Regulation of CD4 T cell reactivity to self and non-self.

While the thymus may be effective in inducing tolerance to lymphoid associated antigens, it is not as efficient in deleting T cells reactive to peripheral tissue specific antigens. Therefore, to maintain self tolerance to peripheral tissues, post-thymic mechanisms must be invoked. One important way to prevent autoimmune pathology mediated by autoreactive CD4 T cells is the diversion of clones to regulatory Th2 effector cells. However, many different factors contribute in vivo to the decision of stimulated CD4 T cells to develop into Th1 versus Th2 cells. For example, T cell signaling pathways may influence the types of cytokines produced by naive T cells, and studies have provided evidence for a genetic polymorphism among common mouse strains that can significantly influence the early cytokine production in stimulated naive CD4 T cells. The allele carried by the BALB/c strain promotes IL-4 production, and consequently provides resistance to autoimmune diabetes in our transgenic mouse model. In addition, antigen presenting cells can influence the development of stimulated CD4 T cells in part through the production of cytokines such as IL-12. The absorption of IL-12 in vivo can permit the expansion of Th2 type effector cells, and this phenomenon will also protect mice from autoimmunity. Finally, the relative potency of various class II positive antigen presenting cell types can influence the development of autoreactive T cells, with dendritic cells apparently being the strongest stimulator of Th1 responses. Consistent with this notion, a relB knockout mouse, which is missing dendritic cells, appears to drive Th2 development even in response to viral infection. In sum, these various influences over the Th1/Th2 decision in vivo may provide new targets for immunotherapy of autoimmune diseases.

A case of neural cell adhesion molecule-positive peripheral T-cell lymphoma associated with human T-cell lymphotrophic virus type 1 showing an unusual involvement of the gastrointestinal tract during the course of the disease

We report a case of neural cell adhesion molecule (NCAM)-positive peripheral T-cell lymphoma (PTCL), pleomorphic, medium and large cell type according to the updated Kiel classification, which was associated with human T-cell lymphotrophic virus type 1 (HTLV-1). The patient showed an unusual involvement of the gastrointestinal tract forming a bulky mass during the course of his disease, and a relatively indolent clinical course for 53 months until death. Phenotypic analysis showed CD2+, CD3−, CD4+, CD5+, CD7+, CD8−, CD16−, CD56+, and CD57− phenotype. Human T-cell lymphotrophic virus type 1 antibody was detected by the enzyme-linked immunosorbent assay. The proviral integration of HTLV-1 and the monoclonal rearrangements of T-cell receptor beta and gamma genes also were confirmed in DNA extract from fresh material. This fact calls attention to NCAM as one of the potential factors playing a role in the localization and behavior of lymphomas even within the category of PTCL associated with HTLV-1, ie, adult T-cell leukemia/lymphoma (ATLL).

Markers of surface mucous cell type human gastric cancer cells: Galactose oxidase‐Schiff reactive mucins, monoclonal antibody SH‐9 reactive mucins and cathepsin E

Cellular differentiation of gastric cancer cells allows the classification of cell type into surface mucous cell, pyloric gland cell, intestinal absorptive cell and goblet cell types by mucin histochemistry and pepsinogen (Pg) immunohistochemistry. Surface mucous cell differentiation of gastric cancers of each histologic type has previously been detected by the galactose oxidase-Schiff (GOS) reaction although this is not always positive in all cases. Mucus granules of surface mucous cells of normal gastric mucosa show an intense reactivity for SH-9 (monoclonal antibody against CA125-bearing antigenic molecule fragments). Cathepsin E is also expressed in the cytoplasm of surface mucous cells, weakly in absorptive cells of duodenal villi and occasionally in pyloric gland cells. Expression of SH-9 reactive mucin and of cathepsin E were therefore investigated as possible additional markers to distinguish between the gastric cancer cell type in 203 primary stomach cancers. SH-9 reactive mucin was found selectively in GOS positive cancer cells of surface mucous cell type and/or cancer cells unclassified by mucin histochemistry. These latter cells were therefore classified into the surface mucous cell category. Cathepsin E was found mainly in cancer cells of the GOS positive surface mucous cell type and occasionally, in intestinal absorptive and pyloric gland cell types. Galactose oxidase-Schiff, SH-9 and cathepsin E reactive or positive cancer cells were found in 145 (71.4%), 151 (74.4%) and 144 (70.9%), respectively, of the 203 primary stomach cancers investigated.

Immunohistochemical localization of recoverin-like Ca2+-binding protein (P23K) in mouse brain.

An immunohistochemical study was carried out to determine the localization of recoverin-like Ca2+-binding protein (P23k) in mouse brain. P23k was found in pyramidal neurons in the cerebral cortex and hippocampus, granule cells in the cerebellum, periglomerular cells in the olfactory bulb, and neuronal cells in the paraventricular nucleus of the hypothalamus and the posterior mamillary nucleus. In most cell types, P23k was localized mainly on the plasma membrane, and partly in the cytosol. Since P23k has a Ca2+-dependent membrane-binding property, the difference in the subcellular localization of P23k seems to be caused by the variety of intracellular Ca2+-levels in the positive cells. The wide range of positive cell types implies that P23k may have a wide range of physiological functions.