Preferential expansion of human umbilical cord blood-derived CD34-positive cells on major histocompatibility complex-matched amnion-derived mesenchymal stem cells

Background

We previously found in a murine hematopoietic system that hematopoietic stem cells show high differentiation and proliferation capacity on bone marrow-derived mesenchymal stem cells/stromal cells (microenvironment) with “self” major histocompatibility complex (MHC).

Design and Methods

We examined whether amnion-derived adherent cells have the characteristics of mesenchymal stem cells, and whether these adherent cells can support the proliferation of umbilical cord blood-derived lineage-negative and CD34-positive cells (Lin^–CD34⁺ cells) obtained from the same fetus to a greater extent than those derived from other fetuses.

Results

Culture-expanded amnion-derived adherent cells expressed mesenchymal stem cell markers and HLA-ABC molecules and could differentiate into osteoblasts, adipocytes and chondrocyte-like cells, indicating that the cells have the characteristics of mesenchymal stem cells. The Lin^–CD34⁺ cells purified from the frozen umbilical cord blood were strongly positive for HLA-ABC, and contained a large number of hematopoietic stem cells. When the Lin^–CD34⁺ cells were cultured on the autologous (MHC-matched) or MHC-mismatched amnion-derived adherent cells in short-term assays (hematopoietic stem cell-proliferation) and long-term culture-initiating cell assays, greater expansion of the Lin^–CD34⁺ cells was observed in the MHC-matched combination than in MHC-mismatched combinations. The concentration of granulocyte-macrophage colony-stimulating factor in the culture supernatants of the long-term culture-initiating cell assays was significantly higher in the MHC-matched combination than in MHC-mismatched combinations.

Conclusions

It is likely that a MHC restriction exists between hematopoietic stem cells and mesenchymal stem cells/stromal cells in the human hematopoietic system and that granulocute-macropage colony-stimulating factor contributes to some extent to the preferential hematopoiesis-supporting ability of the MHC-matched amnion-derived adherent cells.     

A case of superficial angiomyxoma

We report a case of a 19-year-old man who showed a brownish, elastic-soft, multilocular, pedunculated, solid tumor (10 × 6 × 6 cm), with scale crust and erosion, on the medial side of the right thigh. The histopathology of a specimen removed completely revealed a tumor that was located between the middle layer of the dermis and the subcutaneous fatty tissue, was filled with a mucoid material, and was surrounded by a fibrous tissue. The tumor consisted of small thin-walled blood vessels and spindle-shaped or stellate tumor cells without cytological atypia in addition to the mucoid material. Immunohistochemistory revealed that the tumor cells express the vimentin stain. Based on these clinical and histologic findings, we diagnosed the skin condition as superficial angiomyxoma.     

Ultrasonic tissue characterization of photodamaged skin by scanning acoustic microscopy

The aim of this study was to ultrasonically characterize photodamaged skin of the elderly at the microscopic level using scanning acoustic microscopy which showed two-dimentional distribution of sound speed in the skin section. We confirmed that the expression level of the elastin gene was increased in the preauricular skin (photodamaged area), compared with postauricular skin (photo-protected area). The expression level of the procollagen gene was also increased in the preauricular skin compared with postauricular skin. The preauricular skin showed higher sound speed in the papillary dermis (Grenz zone). The site of progressive solar elastosis showed a somewhat sound speed velocity than that of the Grenz zone. Immunohistochemical staining showed conserved deposition of collagen in the Grenz zone even in the more photodamaged preauricular skin. These results suggest that fibrosis in the Grenz zone compensates tissue strength with the progress of solar elastosis. The sound speed analysis of skin will provide important information on heterogeneous mechanical changes in the skin during the process of photoaging.     

Cardiac functional analysis with multi-detector row CT and segmental reconstruction algorithm: comparison with echocardiography, SPECT, and MR imaging

PURPOSE: To evaluate accuracy of cardiac functional analysis with multi-detector row computed tomography (CT) and segmental reconstruction algorithm over a range of heart rates. MATERIALS AND METHODS: Institutional review board approval was obtained. Informed consent was not required. Multi-detector row CT (500-msec rotation time, 8 x 1-mm detector collimation) and magnetic resonance (MR) imaging were performed in 50 patients (28 men, 22 women; age range, 46-84 years; mean age, 67 years). Two-dimensional echocardiography was performed in 41 patients, and electrocardiographically (ECG)-gated single photon emission computed tomography (SPECT) was performed in 27. End-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF), and left ventricular (LV) mass were estimated with multi-detector row CT and compared with values estimated with MR imaging, which served as the reference standard. Additionally, EF values estimated with multi-detector row CT, echocardiography, and SPECT were compared with those estimated with MR imaging. Systemic error and degree of agreement of global functional parameters measured with MR imaging and other modalities were assessed. In a second analysis, linear regression analysis was added. RESULTS: EF estimated with multi-detector row CT agreed and correlated well with EF estimated with MR imaging (bias +/- standard deviation, -1.2% +/- 4.6; r = 0.96). Agreement and correlation were similar for EDV (-0.35 mL +/- 15.2; r = 0.97), ESV (1.1 mL +/- 8.6; r = 0.99), and LV mass (2.5 mL +/- 15.0; r = 0.96). Standard deviation of EF difference between multi-detector row CT and MR imaging was significantly less than that between echocardiography and MR imaging (P < .001) or that between SPECT and MR imaging (P < .001). CONCLUSION: Various LV functional parameters were measured with multi-detector row CT with a segmental approach, and measurements correlated and agreed with those obtained with MR imaging. Moreover, functional analysis with multi-detector row CT was more accurate than that with two-dimensional echocardiography or ECG-gated SPECT.     

A novel human monoclonal antibody,TONO-1, reactive with T-lymphocytic leukemia cells

Mononuclear cells from the peripheral blood of patients with systemic lupus erythematosus (SLE) were transformed with the Epstein-Barr virus (EBV) and the resultant polyclonal B-lymphoblastoid cell lines were tested for antibody activity to membrane antigens of certain T-cell lines. B lymphoblastoid cell lines secreting specific antibodies were fused with (mouse x human) heteromyeloma SHM-D33 cells. Among the large number of hybridomas generated, one which produced a human monoclonal antibody (MAb) TONO-I (1gM, λ) was selected. MAb TONO-I proved to be reactive with 4 human T-cell lines, HPB-MLT, L-MAT, MOLT-3 and MOLT-4F, but not with B-leukemia, Burkitt's lymphoma, myelomonocytic leukemia, erythroleukemia or non-hematopoietic malignant cell lines. MAb TONO-I reacted positively with fresh leukemia cells from 2 of 7 patients with acute T-lymphocytic leukemia, but no reaction was observed in non-T-cell leukemia cases. Normal lymphocytes, monocytes, granulocytes, red blood cells and platelets in the peripheral blood did not demonstrate remarkable binding. Neither thymocytes nor bone-marrow cells from healthy volunteers were reactive. The antigens defined by MAb TONO-I were polypeptides of 57 kDa and 68 kDa. Immunohistological studies revealed no staining of thymocytes in the thymus of a 6-month-old child, but showed epithelial reticular cells and Hassall's corpuscles to stain positively. These results suggest that MAb TONO-I is directed to T-leukemic cells and some components of thymus tissue. © 1995 Wiley-Liss Inc.     

Activation of thymic B cells by signals of CD40 molecules plus interleukin-10

We have previously found that thymic B cells, particularly thymic CD5⁺ B cells, show low responsiveness to the usual B cell stimulants such as lipopolysaccharide or anti-IgM plus interleukin (IL)-4, although they proliferate and produce antibodies after direct interaction with major histocompatibility complex class II-restricted T blasts. These findings raise the possibility that a CD40-CD40 ligand (L) interaction is involved in the activation of thymic B cells. In the present study, we therefore examine this possibility using CD40L-transfected Chinese hamster ovary (CHO) cells or anti-CD40 monoclonal antibody (mAb). When B cells in the spleen and peritoneal cavity were stimulated, they proliferated and produced immunoglobulin (Ig) in the presence of CD40L-CHO cells or anti-CD40 mAb alone. However, another signal delivered by IL-10 in addition to CD40L-CHO cells or anti-CD40 mAb was found to be necessary for thymic B cells to proliferate and secrete Ig. Other interleukins acting on B cells, such as IL-4, IL-5, and IL-6, had no effect on the activation of thymic B cells, which thus have unique characteristics not found in peripheral B cells. This report discusses the physiological significance of IL-10- and CD40-driven signals in the activation of thymic B cells.     

IL-17 and IL-17F modulate GM-CSF production by lung microvascular endothelial cells stimulated with IL-1beta and/or TNF-alpha

In this study, we investigated the roles of CD4 T cell cytokines IL-17 and IL-17F in GM-CSF production from lung microvascular endothelial cells (LMVECs). While a wide range of doses of IL-17 or IL-17F alone did not induce GM-CSF release from LMVECs, IL-17 had an enhancing effect on macrophage-derived IL-1beta- and TNF-alpha-induced GM-CSF mRNA expression and production, whereas IL-17F had an enhancing effect on IL-1beta-induced GM-CSF production, but a marked inhibitory effect on TNF-alpha-induced secretion. GM-CSF production was further enhanced with the combination of three cytokines IL-1beta, TNF-alpha and IL-17 or IL-17F. Additionally, when Th1 or Th2 cytokine was combined with IL-1beta or TNF-alpha, both Th1 and Th2 cytokines had a modest stimulatory effect on TNF-alpha-induced GM-CSF production, whereas IL-4 and IFN-gamma profoundly attenuated IL-1beta-induced secretion. Moreover, the regulation by IL-17 plus Th1 or Th2 cytokine of GM-CSF production from LMVECs treated with IL-1beta or TNF-alpha was dependent on the concentration of IL-17. Our findings indicate that IL-17 and IL-17F play a differential regulatory role in GM-CSF production by LMVECs stimulated with IL-1beta and/or TNF-alpha, which is sensitive to Th1 and Th2 cytokine modulation.     

Anaphylactoid purpura with intestinal perforation: Report of a case and review of the Japanese literature

A 78 year old man suffering from anaphylactoid purpura complained of abdominal pain and bloody stools. Combined therapy with Prednisolone and cyclophosphamide had been given against nephrotic syndrome caused by purpura nephritis. Severe abdominal pain with symptoms and signs of panperitonitis necessitated laparotomy. Rectosigmoid perforation due to necrotizing angiitis (phlebitis) was also demonstrated histologically. The patient died of septicemia 18 days after surgery. Perforation of the alimentary tract rarely occurs in anaphylactoid purpura. Twenty similar cases were briefly reviewed from the Japanese literature.     

Effects of intrathymic injection of organ-specific autoantigens,parietal cells,at the neonatal stage on autoreactive effector and suppressor T cell precursors

Thymectomy on day 3 after birth (3d-Tx) induces autoimmune gastritis (AIG) in 81%, and oophoritis (AIO) in 25% of BALB/c mice at the age of 2 to 3 months. Intrathymic, but not intraperitoneal injection of syngeneic parietal cells into sex-matched BALB/c mice within 24 h of birth resulted in almost complete prevention of the development of AIG in these mice in which 3d-Tx was performed. The prevention induced was parietal cell specific, since the development of AIO was not inhibited in female mice. Moreover, the injection of BALB/c liver cells, Mls-matched (BALB/c) and -disparate (DBA/2) B blasts which resulted in Vβ6 T cell deletion, as well as the injection of staphylococcal enterotoxin B failed to prevent the diseases. These findings suggested that recognition of an autoantigen in the thymus is necessary for the induction of tolerance, and that involvement of Mls-1 antigens in the pathogenesis of AIG, as has been suggested previously (Schwartz, R. H., Cell 1989. 57: 1073), was unlikely. T cells that suppress the development of organ-specific autoimmune diseases in 3d-Tx mice seem to maintain the unresponsiveness of autoreactive T cells at the periphery in normal mice. In agreement with our previous observations, we found that intraperitoneal (i.p.) injection of spleen cells from 3-month-old normal mice into 3d-Tx mice on day 10 after birth prevented the development of AIG, whereas spleen cells from age-matched AIG⁺ (mice with AIG) or AIG^? (mice without AIG) 3d-Tx mice failed to do this. This implies that the suppressor cells probably affect the differentiation of effector-precursor to effector. In fact, these suppressor cells did not inhibit the adoptive transfer of AIG to nu/nu BALB/c mice by spleen cells from 3d-Tx mice manifesting AIG. By negative selection using monoclonal antibody and complement, it was confirmed that the phenotype of the suppressor cell was CD4. In contrast to 3d-Tx, 10d-Tx did not induce AIG, indicating the peripheralization of the suppressor cell by that time. On the other hand, intrathymic injection of parietal cells immediately after birth did not affect suppressor cell generation, implying that some T cells, including suppressor cells, escape thymus selection. We postulate that these cells correspond to the precursors of the autoreactive effector T cells and suppressor T cells that are present in normal mice.