Cell reactions to metastatic tumors in the regional lymph nodes were studied by light and electron microscope in 20 cases; i.e. reactive hyperplasia (3), tuberculosis (3), metastatic carcinomas from the breast (4), from the stomach (2), from the lung (2), metastatic epidermoid carcinoma (2), metastatic malanoma (2), and reticulum cell sarcoma (2). The lymph node response was usually germinal center predominence type and the pyroninophilic cell response was a similar pattern of nonspecific germinal centers with prominent reactive hyperplasia. In two cases of undifferentiated tumors, one from the breast and another from the lung, large numbers of pyroninophilic cells were found within the tumor tissue. However, the majority of lymphoid cells surrounding tumor cell or tumor masses were pyronin negative lymphocytes. Electron microscopic observations revealed that the cells surrounding tumor cells were mostly medium sized lymphocytes, occasionally blast cells and mature plasma cells. The contact border between the tumor cells and the surrounding cells was mostly tight and smooth, but occasionally loose with irregular processes, and widely separated in the case with plasma cells. Degenerative changes of adjacent cytoplasm of either the tumor cells or the lymphocytes were not frequent, but in some instances focal degeneration of adjacent cytoplasm, particularly on the side of the lymphocytes, was noted.
Human ; Lymph Nodes/ultrastructure* ; Lymphatic Metastasis ; Lymphocytes/ultrastructure ; Neoplasm Metastasis ; Neoplasms/ultrastructure* ; Plasma Cells/ultrastructure

Alternations of lymphocyte in biophysical properties (e.g., morphology and viscoelasticity) are related to the human health, disease diagnosis and treatment. Here, we used atomic force microscopy (AFM) to characterize the morphology and mechanical properties of normal lymphocyte and Jurkat. The AFM images revealed that their cell shapes appeared similar. The mechanical properties of the two groups were tracked with AFM-based force spectroscopy. The normal lymphocyte cells had a high adhesion force distribution in (796.7 +/- 248.5) pN, whereas the Jurkat cells had a low force distribution in (158.5 +/- 37.5) pN. The adhesion force revealed that the Young's modulus of normal lymphocyte cells (0.471 kPa +/- 0.081 kPa) was nearly four times higher than that of Jurkat cells (0.0964 kPa +/- 0.0229 kPa) at the same loading rate. The stiffness of normal lymphocyte cells was (2.278 +/- 0.488) mN/m and that of Jurkat cells was (4.322 +/- 0.382) mN/m. The differences in mechanical properties of normal and cancerous cells were obvious that healthy and diseased states could be clearly distinguished. These results may be applied to the clinic disease diagnosis for distinguishing the normal cells from the cancer ones even when they show similar shapes.
Biomechanical Phenomena ; Humans ; Jurkat Cells ; physiology ; ultrastructure ; Lymphocytes ; physiology ; ultrastructure ; Microscopy, Atomic Force ; methods

In order to investigate the ultrastructural features of malignant T cell (MTC) in bona marrow aspirate (BMA) from patients with T Cell Lymphoma, the antigen expression of MTC was analyzed by flow cytometry, and the ultrastructural features of MTC in BMA from 13 T-cell lymphoma patients with bone marrow involvement (BMI) were observed by transmission electron microscopy. The results indicated that the sizes of MTC were uneven in every patient and their diameter were between 12 and 28 microm, in 6 out of 13 cases sizes of MTC were slightly uneven but in 7/13 cases sizes of MTC were significantly uneven. The heterochromatin of MTC was less than that of normal T cell and nucleolus diameter was from 2 to 8 microm in all cases. The nuclear contour of MTC was strikingly irregular in 10 out of 13 cases. The MTC had plenty of cytoplasm in 8 out of 13 cases and displayed many microvilli or processes on MTC surface in 7 out of 13 cases, while MTC in 6 out of 13 cases contained more Golgi's apparatuses, secretary vacuoles, dense granules and intermediate filaments. In 8 out of 13 cases mitochondria apparently swelled. It is concluded that the size of MTC increase unevenly in all patients. MTC nuclear contour in most cases is irregular by folding, indenting, and twisting, which often correlated with arising of paranuclear intermediate filaments. Processes and microvilli on surface and Golgi's apparatus, secretary vesicles, dense granules as well as intermediate filament in cytoplasm of MTC develop synchronously, meanwhile, mitochondria of MTC strikingly swell in most cases.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; ultrastructure ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Lymphoma, T-Cell ; pathology ; ultrastructure ; Male ; Middle Aged ; Neoplasm Invasiveness ; T-Lymphocytes ; ultrastructure

OBJECTIVETo establish a modified method for microculturing whole human blood for cytogenetic analysis.

METHODSA novel tube rack was designed to overcome the drawbacks of directly culturing the cells within centrifuge tubes. The fractions of human plasma, human serum and two commercial fetal bovine sera were analyzed with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The influence of adding 0%, 5%, 10%, 15%, 20%, 25% and 30% autologous plasma to the culture on lymphocyte transformation rate and mitotic index (MI) was examined.

RESULTSThe SDS-PAGE analysis showed a significant difference between commercial fetal bovine sera, and that the components of human plasma were similar to those of fetal bovine serum. The value of MI in lymphocyte was evidently increased along with addition of autologous plasma. However, this has exerted no significant effect on the transformation rate. With the addition of 10% autologous plasma, the MI value has become much higher than the conventional method.

CONCLUSIONA modified method was established by application of a novel tube inclined rack and optimization of whole blood inoculation. This method is easier and cheaper, and is suitable for application in clinical practice.

Adolescent ; Adult ; Cell Culture Techniques ; methods ; Cytogenetics ; Female ; Humans ; Lymphocytes ; ultrastructure ; Male ; Mitotic Index

OBJECTIVETo investigate the changes of relative telomere length (RTL) of peripheral blood (PB) CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺T lymphocytes, CD19⁺B lymphocytes and bone marrow (BM) CD34⁺ cells and its association with disease severity in untreated patients with immuno-related pancytopenia (IRP).

METHODSThe PB CD3⁺ , CD3⁺ CD4⁺ , CD3⁺ CD8⁺ T lymphocytes, CD19⁺ B lymphocytes, and BM CD34⁺ cells were purified by magnetic activated cell sorting (MACS), and RTL were measured with flow-fluorescence in situ hybridization (FLOW-FISH).

RESULTSThe RTL of CD3⁺, CD3⁺CD4⁺ , and CD3⁺CD8⁺T lymphocytes in untreated IRP patients were (27.754 ± 16.323)%, (7.526 ± 3.745)% and (25.854 ± 14.789)%, respectivly, which were significantly shorter than those in healthy-controls (54.555 ± 19.782)%, (12.096 ± 2.805)%, and (38.367 ± 4.626)% (P<0.05). The RTL of CD19⁺ lymphocytes in untreated IRP patients was (22.136 ± 16.142)%, which was significantly shorter than that in healthy controls (42.846 ± 16.353)% (P<0.01). There was no significant difference of BM CD34⁺ cells RTL between the untreated IRP patients (22.528 ± 21.601)% and the healthy controls (23.936 ± 19.822)% (P>0.05). There were significantly positive correlations between the RTL of B lymphocytes and the count of white blood cell (r=0.706, P=0.015). There were negative correlations between RTL of B lymphocytes and the clinical symptoms (r=-0.613, P=0.045) and positive correlations with therapeutic effect (r=0.775, P=0.005).

CONCLUSIONThe shorter RTL of CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, CD19⁺ lymphocytes, and the normal RTL of BM CD34⁺ cells in untreated IRP patients were identified, which might imply that IRP is a type of acquired autoimmune diseases.

Adolescent ; Adult ; B-Lymphocyte Subsets ; immunology ; Child ; Female ; Humans ; Lymphocytes ; ultrastructure ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology ; T-Lymphocyte Subsets ; immunology ; Telomere ; ultrastructure ; Young Adult

OBJECTIVETo identify clinical and pathological features of giant cell myocarditis.

METHODSClinical presentation and follow-up data of three patients with giant cell myocarditis were collected.Gross, histopathological, immunohistological and ultrastructural findings of extransplantated hearts of the patients were documented.

RESULTSGrossly, multifocal involvement of the myocardium with variably dilated cardiac chambers were observed in all 3 cases.Histological examination revealed pronounced focal inflammatory infiltrates with multinucleated giant cells. Multinucleated giant cells were positive for CD68 and CD11b immunostains but were negative for CD163 in all cases. Transmission electron microscopy showed that the multinucleated giant cells derived from fusion of several macrophages with adherent lymphocytes and secretary cells. Clinically, the overall patient condition improved in all three cases after heart transplantation.One patient experienced acute cellular rejection (2R level) 4 months after transplantation, but recovered after treatment. One patient developed multinucleated giant cells observed in heart biopsy two weeks after transplantation.

CONCLUSIONSGiant-cell myocarditis is a rare disease of adult, and cardiac transplantation could improve the clinical outcome. Multinucleated giant cell in the myocarditis lesions were derived from macrophages, likely participating in the immune response. Endomyocardial biopsy is important for the diagnosis of giant cell myocarditis.

Acute Disease ; Adult ; Biopsy ; Giant Cells ; pathology ; ultrastructure ; Heart Transplantation ; Humans ; Lymphocytes ; pathology ; Macrophages ; pathology ; Microscopy, Electron, Transmission ; Myocarditis ; pathology ; Myocardium ; pathology ; ultrastructure

Atomic force microscope (AFM) was used to study biotoxicity of food preservative sodium benzoate (SB) at the single cellular level. Lymphocyte morphology and membrane ultrastructure treated with SB at different concentrations and time were analyzed visually. As compared to the normal lymphocyte, the cell morphology and membrane was significantly changed and its ultrastructure was also complicated. After treated with SB, the Rp-v, Rq, Ra and Z values were changed. The statistical analysis of lymphocytes after treated with SB was studied, and discussed its mechanism.
Animals ; Cell Membrane ; ultrastructure ; Lymphocytes ; drug effects ; pathology ; Mice ; Mice, Inbred BALB C ; Microscopy, Atomic Force ; Sodium Benzoate ; toxicity

Single cell gel electrophoresis assay (SCGE), also named as alkaline comet assay, was a simple, rapid and sensitive method to evaluate DNA damage. In this study SCGE technique was used to monitor DNA damage difference in tumor patients caused by chemotherapy, DNA damage distribution frequency and DNA damage characters were analyzed by komet image analysis system (KIAS). The results showed that cyclophosphamide greatly caused DNA damage in lymphocytes of tumor patients. There was significant difference of peripheral blood lymphocyte DNA damage between tumor patients and healthy controls. Tail length of lymphocytes were 33.69 +/- 7.56 micro m, and tail DNA% we re 31.51 +/- 5.4 6% in 10 cancer patients treated with cyclophosphamide, while Tail length were 1 6.2 +/- 1.5 micro m and tail DNA% were 7.46 +/- 1.15% in healthy controls. there was great significant difference on tail length and tail DNA% values between cancer patients and healthy controls (P < 0.01). In conclusion, the successful measurement of DNA damage caused by Cyclophosphamide treatment means that the alkaline comet assay as a valuable tool can be very useful in cancer epideminology study, and also be valuable to evaluate DNA damage status of patients in clinic.
Comet Assay ; Cyclophosphamide ; adverse effects ; DNA Damage ; Electrophoresis, Agar Gel ; Humans ; Lymphocytes ; drug effects ; ultrastructure ; Neoplasms ; drug therapy ; genetics

OBJECTIVETo explore whether there is difference in arsenicals-induced DNA damage of human lymphocyte.

METHODSLymphocyte were sterilely collected from healthy donor and exposed to sodium arsenite (AsIII), sodium arsenate(AsV) and methyl sodium arsenate(MAsv) at 1,5,10,20 and 50 mumol/L. After incubation of 24 hours, cells were collected by centrifugation and DNA damage was detected by single cell gel electrophoresis (SCGE).

RESULTSThe comet frequency distribution of all groups except 1 mumol/L group of MAsV were significantly different from that of control. The comet length of all groups except 1 mumol/L group of AsV and 1.5 mumol/L groups of MAsV were significantly higher than that of control. There were correlations between the doses of arsenicals and the ratios of comet cell or length of comet(rAsIII = 0.8134, rAsV = 0.8734, rMAsV = 0.8994).

CONCLUSIONDNA damage in human lymphocyte were induced by all the three arsenicals. A dose-effect relationship was observed between exposure doses of the same arsenical and DNA damage. With different arsenicals but the same exposure dose, the DNA damage level was as follow: AsIII > AsV > MAsV.

Arsenates ; toxicity ; Arsenites ; toxicity ; Comet Assay ; DNA Damage ; Dose-Response Relationship, Drug ; Humans ; Lymphocytes ; drug effects ; ultrastructure

OBJECTIVETo isolate exosomes from rabbit aqueous humor and to investigate their immunosuppression function.

METHODSAqueous humor was collected from 40 New Zealand rabbits and exosomes were isolated by fractional separation and ultracentrifugation methods; the morphology was studied with electron microscopy. The immunosuppressive-related proteins of exosomes were detected with Western blotting; their inhibitory effect on ConA-induced proliferation of T lymphocyte was estimation with CCK-8 cells proliferation assay.

RESULTSEight milliliters of aqueous humor were collected from 40 New Zealand rabbits and 200 μg exosomes was yielded. Under electron microscope, the exosomes had typical structure of lipid bi-layer with a diameter of 50-100 nm. The results of Western blotting showed that these exosomes expressed Hsp70, CD9 and Alix but not Grp94, presenting a typical exosomes protein profile. Moreover, exosomes expressed high level of TGF-β and significantly inhibited the proliferation of T lymphocytes.

CONCLUSIONImmunosuppressive exosomes can be isolated from rabbit aqueous humor, which may be involved in immunotolerance of the eye.

Animals ; Aqueous Humor ; immunology ; Cells, Cultured ; Exosomes ; immunology ; metabolism ; ultrastructure ; Female ; Immune Tolerance ; Male ; Rabbits ; T-Lymphocytes ; immunology