Large Numbers Of Lymphoid Cells Research Articles

Pathomorphological changes in parenchymatose organs of rabbits in case of chronic passalurosis

Pathomorphological changes, developing in case of pasalurosis chronic course in rabbits are described in the article. The research was held on ten rabbits of Large Grey breed, which were held on household farms of Poltava region. The diagnosing was complex: based on anamnesis data, clinical symptoms’ analysis, and the results of pathomorphological examination. The fragments of the liver, spleen, and kidneys were taken for histological analysis. While making histological analysis of the spleen it was revealed that lymphoid nodules were enlarged, some of them were of uniform structure, and others were in the state of hyperplasia. Non-uniformity of structure in addition to changing the size was registered in those nodules: there was less concentration of lymphoid cells in their central part and more concentration on the periphery. The situation was contrary in separate lymphoid nodules. The red pulp consisted of a large number of lymphoid cells and erythrocytes, the largest number of which were registered in the zones adjoining the places of lymphoid nodules’ location. Sometimes there were megakaryocytes in the field of vision. It was established, that blood filling of arteries was less than normal. In case of using some preparations, bilirubin inclusions were found at having big enlargements in red pulp. Grainy and fatty hepatocytes’ dystrophies, and also lymphohistiocytic interstitial hepatitis were also registered. At the same time, renal corpuscles were enlarged. Rather big spaces, filled with transparent substance containing vessel and cellular fragments (detritus), were revealed in the majority of them between glomeruli and the capsule.Capsule epithelial cells were destroyed in some places. Petechial hemorrhages were registered inside glomeruli. The destruction of separate podocytes and deep cells was noticed. The tubular epithelial cells were enlarged in size, and in some cases they were of cylinder, but not cubic shape; the spaces of such tubular were becoming correspondingly less. The epithelial cell cytoplasm of the tubular in some cases was not uniformand it was of cloudy-grey color: in other cases it had rather large colorless areas and, probably, looked like lipid inclusions (which were washed out from the cells as a result solvent impact during making histological preparations). Concentrations of eosinophilic masses (protein cylinders) sometimes containing certain amount of erythrocytes were often found in canaliculi spaces. Sometimes the destruction of epithelial cells was registered in the tubulеs, and their fragments were found in spaces. As a result of histological examination, the diffuse lymphohistiocytic infiltration in the renal interstitial tissue was found.

Histology and scanning electron microscopy of the tubal tonsil of goats.

Aim:To observe the light and scanning electron microscopy (SEM) of the caprine tubal tonsil.Materials and Methods:The study was conducted on six crossbred male goats of 6 months of age. From the median sections of the head, tissue pieces from the nasopharynx around the auditory tube were collected and fixed for histology and SEM.Results:Tonsillar lymphoid tissue was located in the nasopharynx ventral to the auditory tube opening in the lateral wall of the pharynx. The height of the surface epithelium of the tubal tonsil measured 80.17±1.08 µm and was a pseudostratified ciliated columnar type with basal, supporting, and goblet cells. Above the dome of lymphoid nodules, the epithelium was modified into a follicle associated epithelium (FAE), also called lympho-epithelium or reticular epithelium and was characterized by the absence of goblet cells and cilia, reduced number of cell layers, and a large number of lymphoid cells due to interrupted basement membrane. The height of FAE was smaller than that of the surface epithelium and measured 34.33±0.92 µm. The surface of tubal tonsil showed folds and invaginations, which formed crypts. The lamina propria-submucosa underneath the epithelium was formed by the meshwork of reticular and, thin and loose collagen fibers with dome-like accumulation of lymphoid nodules. In the secondary lymphoid nodules, a corona, parafollicular area, and interfnodular area were observed. The average number of lymphoid nodules counted per field under low power magnification of microscope was 1.17±0.17, and the internodular distance was 34.00±4.37 µm. The mean diameter of lymphoid nodules was 566.67±11.45 µm and the lymphocyte count per nodule was 14741.67±174.36. The number of plasma cells counted per field under low power was 44.38±2.90 below the surface epithelium. The tubal tonsil was not encapsulated. In SEM, the surface epithelium of the tubal tonsils presented ciliated cells, microvillus (MV) cells, and goblet cells. The region of FAE possessed Type-I and Type-II MV cells and microfold (M) cells in between.Conclusion:It was concluded that the tubal tonsils were well developed in goats, which might serve as a means of protection against the spread of infection to the middle ear cavity.

Primary cutaneous blastic plasmacytoid dendritic cell neoplasm: two case reports

Objective To report two cases of blastic plasmacytoid dendritic cell neoplasm(BPDCN). Methods The clinical and histopathological manifestation, immunophenotype, treatment and prognosis of BPDCN were analyzed in two patients. Results Both of the patients were female with skin lesions as the initial manifestation. Case 1 presented with purple nodules on the waist, back and left upper limb; histopathological examination revealed that a large number of lymphoid cells diffusely infiltrated the dermis and subcutaneous fat layer, with an uninvolved zone separating the infiltrate from the epidermis. Case 2 presented with a subcutaneous mass around the left knee joint, and histopathological examination showed nodular lymphoid infiltrates in the dermis and subcutaneous tissue. Immunohistochemistry showed positive staining for CD4, CD56, CD123(strong)and Ki-67(40%), but negative staining for myeloperoxidase and EBER in both cases, and positive staining for TdT in case 1 but not in case 2. A bone marrow biopsy disclosed active hyperplasia of granulocytic cells, erythroid cells and megakaryocytes, and increased proportion of segmented cells. Immunophenotype analysis revealed that bone marrow was unaffected. Blood routine examination showed anemia in both patients. Patient 1 died two months after the final diagnosis. After chemotherapy with VDLP(vincristine, daunorubicin, L-asparagine and dexamethasone)and CHOP(cyclophosphamide, doxorubicin, vincristine, and prednisolone), skin lesions subsided in patient 2 who also received allogeneic bone marrow transplantation. At the time of this writing, patient 2 was in good condition with no recurrence of skin lesions. Conclusions BPDCN is a rare malignant neoplasm characterized by rapid growth and strong invasion capacity, usually with skin lesions as the initial manifestation. The prognosis of BPDCN is poor and there is no standard treatment. Early diagnosis and timely treatment may improve the prognosis of BPDCN, and allogeneic bone marrow transplantation may markedly extend survival. Key words: Skin neoplasms; Dendritic cells; Stem cells; Treatment outcome; Prognosis

Progressive Diplopia and Facial Weakness in a 62-Year-Old Woman

Progressive Diplopia and Facial Weakness in a 62-Year-Old Woman

Regulatory T cells contribute to the immunoregulatory effect on graft versus host reaction after liver transplantation in donor-dominant one-way MHC matching rats

Acute graft-versus-host disease following liver transplantation (LTx-aGVHD) poses a major diagnostic and therapeutic challenge, and the mortality rate is as high as 85%. Even the liver contains large numbers of lymphoid cells in the parenchyma and surgical damage of liver transplantation would induce an immunosuppressive, the incidence of LTx-aGVHD is just approximately 1–2%. CD4 +CD25 +regulatory T (Treg) cells have recently been shown to suppress proliferative responses of CD4 +CD25 −T cells to alloantigenic stimulation in vitro and are required for ex vivo tolerization of donor T cells, which results in their reduced potential to induce aGVHD after bone marrow transplantation. To investigate the role of Treg in LTx-aGVHD, we compared the proportional frequency of Treg in syngeneic liver transplantation recipients (sLT group), semiallogeneic liver transplantation recipients (Semi-LT group), LTx-aGVHD induced recipients (LTx-aGVHD group) and healthy controls. Here we show that replacement of (LewisXBN)F1 liver by Lewis liver alone in Semi-LT group was not sufficient to induce aGVHD, and all recipients grew in a normal pattern as the syngeneic LT from Lewis to Lewis rat. However, when 4 × 10 8 of donor splenocytes were transferred simultaneously with LT, the morbidity of lethal aGVHD were 100%. A relative stable percentage of Treg, defined as CD4, CD25 and Foxp3, was detected in peripheral blood mononuclear cells (PBMCs) of sLT group compared with healthy controls. In early time after transplantation, no significantly change of Treg population was observed in these recipients after semiallogeneic liver transplantation in comparison with healthy controls ( P > 0.05). However, Treg levels showed a relative increase 4 days after transplantation. Especially on 12th and 16th day after transplantation, there was significantly increased proportion of Treg cells compared with healthy controls ( P < 0.05). The present of Treg decreased progressively in LTx-aGVHD group, which was significantly lower than group 1 and group 2 on the 12th and 16th day after liver transplantation ( p < 0.05). In conclusion, recipients in semi-LT group harbor an increased percentage of Treg in peripheral blood compared with controls. Treg have an immunoregulatory effect on graft versus host reaction after liver transplantation. Additional donor splenocyte transplanted with the liver provoke the development of aGVHD after liver transplantation, and reversed the change of Treg in PBMCs as in Semi-LT group, which destroy the balance between Treg and conventional effector T cells which determine the outcome of aGVHD after liver transplantation.

In vivoboosting of lung natural killer and lymphokine-activated killer cell activity by interleukin-2: comparison of systemic, intrapleural and inhalation routes

Natural killer (NK) cells are thought to play a role in host defence against malignancy and infection, in immunoregulation and as precursor cells in a generation of lymphokine-activated killer (LAK) cells which can lyse NK-resistant tumour cells. As the lung is a major site for malignancy and infection and as there are large numbers of lymphoid cells including NK cells in the interstitial compartment of the lung, we evaluated the capacity of interleukin-2 (IL-2), a lymphokine capable of augmenting NK activity in vitro, to augment lung NK cell activity in vivo, using different routes of IL-2 administration. We compared both systemic (i.v. and i.p.) and local (intrapleural and inhalation) routes of IL-2 administration (50,000 U/daily for 5 days) using CBA mice, assessing NK and LAK cell activity in the spleen (systemic) and in the lung. The target cells used for these studies were the YAC-1 (NK-sensitive) and P815, NO36 and HA56 (NK-resistant, LAK-sensitive) cell lines. Splenic NK activity was increased by 1.4-1.9-fold for i.v./i.p., respectively, compared with controls with both systemic routes of administration, and lung NK activity was increased 3.2-fold and 3.8-fold (i.v./i.p, respectively, P less than 0.05), to levels which were comparable to systemic (splenic) NK activity following the same therapy. Intrapleural IL-2 administration similarly enhanced lung NK activity (3.3-fold) and splenic NK activity (1.3-fold; P less than 0.05 versus controls for both). Surprisingly, inhaled IL-2 suppressed both splenic and lung NK cell activity (84 +/- 8% and 78 +/- 10% suppression, respectively, P less than 0.05). LAK cell activity was also enhanced in the lung by 1.8-8-fold in response to i.v., i.p. and intrapleural IL-2, whereas inhaled IL-2 was ineffective in generating LAK cell activity. These results suggest that the systemic and intrapleural administration of IL-2 effectively boost pulmonary NK and LAK activity whereas inhalation of IL-2 does not. Thus, in clinical situations where boosting of local lung NK or LAK cell activity is desired, these routes of IL-2 administration may be effective.

Küttner's tumor of the submandibular glands: Report of five cases with fine‐needle aspiration cytology

Küttner's tumor (KT) is a benign tumor-like lesion of the salivary gland that mimics neoplasm clinically because of presentation as a hard mass. Recently, the histomorphological and immunohistochemical findings of this lesion have been analyzed, and differential diagnostic problems relating to salivary gland lymphoma have been discussed. However, currently there is little information on the cytological findings of those lesions. We present cytological findings from five such cases using fine-needle aspiration cytology (FNAC). FNAC of this lesion may present a diagnostic challenge to the cytologist as lesions share some cytologic features with inflammatory process containing numerous lymphoid cells. Smears obtained from two cases contained moderate to large numbers of lymphoid cells without definite cytological atypia, scattered ductal structures, and acinar cell clusters. The remaining three cases showed low cellularity probably attributable to fibrosis that made it difficult to aspirate the cellular element. FNAC findings of scattered ductal structures surrounded by collagens and infiltrated by a mixed population of lymphoid cells, not specific for KT, are highly suggestive of the diagnosis with the appropriate clinical findings. However, a portion of cytological specimens of KT containing relatively large numbers of lymphoid cells should be differentiated from malignant lymphoma arising from the submandibular gland.

Differences in kinetics of donor lymphoid cells in response to G-CSF administration may affect the incidence and severity of acute GvHD in respective HLA-identical sibling recipients.

Granulocyte-colony stimulating factor (G-CSF), in addition to myeloid and stem cells, mobilizes a large number of lymphoid cells. We examined which lymphoid populations were mobilized in 21 consecutive donors of peripheral blood stem cells (PBSC) and whether the differences in mobilization could affect the incidence of acute and chronic GvHD in respective HLA-identical recipients. G-CSF administration induced significant increases of donor B (CD3-CD19+) lymphocytes and slight increases of T (CD3+) and cytotoxic (CD16+CD56+) NK cells. The number of extrathymic cells (CD3+ cells with NK markers, or CD7+) remained unchanged except for an increase of CD3+CD57+CD8+ cells. Donors of patients without subsequent grade II-IV acute GvHD compared to donors of patients who developed significant acute GvHD were found to have in peripheral blood stable numbers of CD3+CD4+ cells producing IL2, with a concomitant increased number of CD3+CD4low+CD25+ T regulatory cells and decreased NK-mediated cytotoxicity, together with a higher number of suppressive extrathymic CD57+CD3+ cells in the blood and G-PBMC grafts. Increasing numbers of activated T and NK cells in the blood were associated with the development of chronic GvHD. We suggest that differences in steady-state levels and kinetics of G-CSF induced mobilization of donor lymphoid cells may in addition to other well-known factors affect the incidence of GvHD in HLA-identical recipients. However, owing to the small number of donor-recipient pairs studied, our results must be verified in a larger group of patients.

Distribution of cell adhesion molecules on CD56++, CD3-, CD16- large granular lymphocytes and endothelial cells in first-trimester human decidua.

Human decidua exhibits a unique infiltrate of large granular lymphocytes (LGL) with a natural killer (NK) cell phenotype (CD56++, CD16-, CD3-). The mechanisms underlying the binding of circulating LGL to vascular endothelium in the decidua and their migration into the decidual stroma were investigated immunohistochemically in first-trimester decidua with antibodies against endothelial adhesion molecules and their counter-receptors on leukocytes. Decidual and peripheral blood LGL were also investigated by flow cytometry. In the immunohistochemical investigations, moderate to large numbers of lymphoid cells in the decidua were found to express the alpha 4 and alpha L integrin subunits, platelet endothelial cell adhesion molecule (PECAM) and intercellular adhesion molecule-1 (ICAM-1). PECAM and ICAM-1 were found on the endothelium of large numbers of decidual blood vessels of all types. Vascular cell adhesion molecule (VCAM), however, was found on the endothelium of only small to moderate numbers of arterioles and venules and a few capillaries, the latter being the main site of migration of leukocytes into the stroma. Weak staining for endothelial leukocyte adhesion molecule (ELAM) was seen only in a moderate number of blood vessels. Flow cytometry revealed expression of the alpha L integrin subunit by 72 +/- 10% and 97 +/- 3% of decidual and peripheral blood CD56+ LGL, respectively, of the alpha 4 integrin subunit by 85 +/- 7% and 90 +/- 5%, of PECAM by 40 +/- 12% and 30 +/- 15%, and of ICAM-1 by 22 +/- 10% and 1 +/- 1%.(ABSTRACT TRUNCATED AT 250 WORDS)

Formation of lymphoepithelial tissue in the sheep's palatine tonsil.

Formation of lymphoepithelial tissue was studied in 1-, 10- and 21-day-old sheep. From each of the animals one of the tonsils was fixed in 4% glutaraldehyde for light- and electron microscopy while the other was frozen in liquid nitrogen for immunohistochemistry. These examinations revealed sequential histological events during the formation of reticular epithelium. (i) Appearance of a distinct epithelial cell indicates the initiation of the reticulation. The electron density of these epithelial cells is much lower than that of the common keratinocytes but the presence of cytoplasmic tonofibrils and desmosomes provides evidence that they are of epithelial origin. They may represent the precursors of M cells. Their appearance may be followed by expression of Ia+ molecules on the surrounding keratinocytes showed by isolated Ia+ areas in the epithelium. (ii) In the mesenchyme underneath the Ia+ epithelial areas, Ia+ dendritic-like cells emerge which immigrate through the basement membrane into the epithelium establishing a provisional dendro-epithelial tissue. In this stage of the reticular epithelium's formation large Ia+ areas are shown by immunostaining, which include the epithelium and mesenchyme. The origin of the Ia+ dendritic-like cells is uncertain but their distribution and dense accumulation underneath the epithelium suggest that they are transformed tonsillar mesenchymal cells. Similar cell transformation of mesenchymal cells takes place in the bursa of Fabricius prior to development of lymphoepithelial tissue. (iii) The M cell precursors together with the Ia+ dendritic-like cells adapt the epithelium to be suitable for receiving a large number of lymphoid cells. Immigration of the lymphoid cells into the epithelium transforms the dendro-epithelial tissue to a real lymphoepithelial one.

Abelson murine leukemia virus variants with increased oncogenic potential.

A number of strains of Abelson murine leukemia virus (A-MuLV) with various abilities to transform cells have been identified. Among these is the A-MuLV-P90 strain, a mutant derived from A-MuLV-P120 that encodes an A-MuLV protein missing sequences that are normally present at the extreme carboxy terminus of P120 (N. Rosenberg and O. N. Witte, J. Virol. 33:340-348, 1980). This virus transforms NIH 3T3 cells efficiently but does not transform a high frequency of lymphoid cells in vitro or in vivo. In this communication, we show that of the relatively few tumors induced by A-MuLV-P90 nearly all contained new variant viruses that stably expressed either larger or smaller A-MuLV proteins. Strains that expressed larger A-MuLV proteins behaved like A-MuLV-P120 in transformation assays, whereas those expressing smaller A-MuLV proteins induced a high frequency of tumors after a short latent period in vivo but failed to transform large numbers of lymphoid cells in vitro. Thus, these latter viruses separated the requirements for in vitro transformation of lymphoid cells from those for tumor induction. All of the variants differed from A-MuLV-P90 in the carboxy-terminal region of the A-MuLV protein, suggesting that sequences in this region play a key role in the ability of the virus to interact with hematopoietic cells in vivo and in vitro.

Immunohistologic studies in pityriasis rosea. Evidence for cellular immune reaction in the lesional epidermis.

Biopsy specimens of the skin lesions of pityriasis rosea obtained from 15 patients were studied. Characteristic histologic changes were composed of focal intercellular edema, epidermotropism of mononuclear cells often associated with the formation of focal intraepidermal collections of mononuclear cells, and of a perivascular lymphohistiocytic cell infiltration in the superficial dermis. Immunologic analysis using monoclonal antibodies showed that large numbers of lymphoid cells in the perivascular infiltrate reacted with anti-pan-T-cell, anti-helper-inducer subset, and anti-HLA-DR monoclonal antibodies, while the epidermotropic mononuclear cells consisted of helper-inducer cells or suppressor-cytotoxic cells without any predominance pattern. In addition to epidermal Langerhans' cells, some of the dermal infiltrating cells were reactive with monoclonal antibody OKT6. Moreover, there was localized expression of HLA-DR antigen on the keratinocytes. We think that cellular immune reactions are taking place in the lesional epidermis of pityriasis rosea.

Immunocytochemical staining of T and B lymphocytes in serous effusions.

Cell smears from serous effusions containing large numbers of lymphoid cells were stained by the alkaline phosphatase-anti-alkaline phosphatase technique with a panel of monoclonal antibodies, including anti-B and anti-T cell antibodies and anti-HLA-DR. Samples from 17 patients with lymphoproliferative disorders--such as chronic lymphocytic leukaemia and non-Hodgkin's lymphoma--and from 19 patients who had no evidence of lymphoid neoplasia--for example, cases of carcinoma, cardiac failure--were investigated. The majority of lymphoid cells in reactive effusions were T cells, which lacked HLA-DR and showed a marked excess of helper/inducer cells (mean helper to suppressor ratio of 3 X 5). In contrast, lymphoid cells in samples from nine cases of B cell neoplasia were positive for B cell antigen and HLA-DR. In a further four B cell neoplasms most lymphoid cells were reactive T cells. Two cases of T cell lymphoid leukaemia could also be characterised by immunocytochemical staining, both being classified as T helper cell neoplasms. Labelling was performed on routinely prepared, air dried cell smears, which could be stored in the unfixed state for long periods before staining. The technique may therefore be of use in many clinical cytology laboratories for the diagnosis of effusions containing numerous lymphoid cells.

Immunohistologic Studies in Pityriasis Rosea

• Biopsy specimens of the skin lesions of pityriasis rosea obtained from 15 patients were studied. Characteristic histologic changes were composed of focal intercellular edema, epidermotropism of mononuclear cells often associated with the formation of focal intraepidermal collections of mononuclear cells, and of a perivascular lymphohistiocytic cell infiltration in the superficial dermis. Immunologic analysis using monoclonal antibodies showed that large numbers of lymphoid cells in the perivascular infiltrate reacted with anti—pan-T-cell, anti— helper-inducer subset, and anti-HLA-DR monoclonal antibodies, while the epidermotropic mononuclear cells consisted of helper-inducer cells or supressor-cytotoxic cells without any predominance pattern. In addition to epidermal Langerhans' cells, some of the dermal infiltrating cells were reactive with monoclonal antibody OKT6. Moreover, there was localized expression of HLA-DR antigen on the keratinocytes. We think that cellular immune reactions are taking place in the lesional epidermis of pityriasis rosea. (<i>Arch Dermatol</i>1985;121:761-765)

T-Cell Subsets in Cutaneous Sarcoidosis

Skin lesions from four patients with systemic and cutaneous sarcoidosis were studied, by the use of monoclonal antibodies, for the presence of T cells and T-cell subsets. Large numbers of lymphoid cells reacting with anti-pan T-cell (LEU-1) and anti-helper and inducer subset (LEU-3) monoclonal antibodies were observed around and within the sarcoid granulomas in three of the four patients. Only rare LEU-2-reactive suppressor cells were observed in all four patients. Activated T lymphocytes with focal acid phosphatase activity, together with epithelioid cells and multinucleated giant cells with strong diffuse activity of acid phosphatase and nonspecific esterase, were identified within the granulomas. The two patients with active disease demonstrated substantially more T cells in the sarcoid granulomas than did the two patients with chronic disease. Our study results suggest the importance of helper T cells in the formation of the sarcoid granuloma by mononuclear phagocytes and imply that the activity and duration of disease may be related to the T-cell populations.

Plasma Cell Infiltrate of the Primary Tumor as a Source of Early Systemic Protection Against Metastases in Mice23

Histologic examination of sc implants of the syngeneic C3H/He mammary carcinoma MC2 showed that the accumulation of a large number of lymphoid cells in the stroma around the implants in C3H/He mice was a constant feature of the early primary host response. This stromal reaction had two main components, the first composed of small lymphocytes and the second composed of larger blast cells with a high proportion of plasma cells. The surgical removal of a 6-day-old tumor implant with its surrounding stromal reaction, if performed 4 days before systemic dissemination of MC2 cells via injection into the left ventricle, resulted in a more frequent growth of the disseminated cells as compared to the frequency of growth in mice carrying their tumor implants. This increased frequency of growth in surgically cured mice could be reduced by repeated transfusions of plasma from MC2-bearing hosts. Repeated ip injections of antigen, in the form of 2 X 10(5) inactivated tumor cells after tumor excision, did not support the development of strong systemic resistance against metastases. When sublethal whole-body radiation was given before the sc tumor implant, the stromal reaction was much reduced, and the surgical removal of sc tumor implants from irradiated mice resulted in only a minor increase in the growth of tumor cells injected into the circulation compared to the growth of tumor cells injected into irradiated mice carrying their tumor implants. It appears that a strong, local primary immune reaction may act as a temporary accessory lymphoid organ, constituting an early and potent source of systemic immune protective factors.

Cell proliferation in follicle-associated epithelium of mouse Peyer's patch.

Follicle-associated epithelium (FAE) from the mouse Peyer's patch consists of a dome part containing goblet as well as columnar epithelial cells and a crypt part, containing Paneth as well as goblet and undifferentiated epithelial cells. The proportions of columnar epithelial, goblet and Paneth cells in the FAE are similar to those found in normal crypt/villus complexes. Mitosis is confined to the crypts of the FAE. The cell-renewal time for FAE columnar epithelial cells, estimated from experiments using tritiated thymidine, is approximately 60 hours. Large numbers of lymphoid cells are also present in the FAE, particularly in regions distal to the crypts. The similarity between follicle-associated and villus epithelial structure is discussed, together with its possible relevance to "M" cell formation within the FAE.

Separation of functional subpopulations of murine and human lymphoid cells on colloidal silica density gradients. I. Preparation of the Ludox AM ® gradient material and characterization of separation capacities

This report describes a unique modification of an isopycnic density gradient system utilizing as a separating menstrum colloidal silica (Ludox AM ®). The primary advantages of this preparation are: (1) It is chemically defined, allowing extremely reproducible cell separation employing different lots of material; (2) the physical parameters (pH, density, salt concentration) of the final gradient suspension can be manipulated over a wide range of values, allowing for the separation of many different biological materials; (3) it allows separation of very large numbers of lymphoid cells with greater than 95% recovery of applied cells; (4) separated cellular subpopulations can be easily washed free of silica and cellular function is retained. This paper is a report of the preparation and functional characteristics of the gradient material as it relates to the separation of very large numbers of lymphoid cell subpopulations in both mouse and man. Subpopulations of murine and human lymphocytes separated by this gradient material were assayed for IgM synthesis, T-cell mediated cytotoxicity, and lymphokine production.

Immunomorphological lymph node changes in patients with operable bronchogenic squamous cell carcinoma.

Histological changes in tracheobronchial and bronchopulmonary lymph nodes of patients with operable (i.e. stages T1/2, N0/1, M0) bronchogenic squamous cell carcinoma were examined histometrically. Out of 29 patients, 10 survived less than 12 months and 19 lived longer than two years after surgery. Quantitative methods were employed for measuring structural changes in lymph node sections at the histological and cellular level. The following parameters reflecting both cellular and humoral immune responses correlated with survival: relative numbers of large lymphoid cells and mitotic figures in the paracortical ("thymus-dependent") area; volume of the follicular (predominantly B-cell) cortex; and volume of germinal centers. Pronounced accumulation of histiocytes or dust-loaded macrophages in the paracortex and presence of "empty" lymph sinusoids (i.e. without sinuhistiocytosis) correlated inversely with survival. In addition to the theoretical importance and considerations of a documented immune response, such data could help in identifying high risk groups within the same stage of bronchogenic carcinoma of a given type.

Studies on lymphocytes: XV. Analysis of the in vivo division cycle of large lymphoid cells in calf thoracic duct using combined microspectrophotometry and autoradiography

From previously reported labeled mitosis curves, the majority of proliferating large lymphoid cells in the thoracic duct of the calf have a T G of about 5 h. The division cycle in vivo of these elements was further investigated by combining microspectrophotometry and autoradiography of a sample taken 20 min following i. v. injection of 3H-thymidine. Compared with small lymphocytes, unlabeled large lymphoid cells with a DNA content near the 2 c mode showed approx. 4% higher mean, and a greater coefficient of variation of, Feulgen absorbance values per nucleus. In contrast to small lymphocytes, they exhibited a frequency distribution with regard to DNA content which diverged from normality with skewness to the right. Large lymphoid cells with a relative DNA content of 1.9–2.3 c showed a significantly smaller mean nuclear surface area in smears than elements with a relative DNA content of 3.7-3.1 c although there was a considerable overlap between the two groups of cells with regard to size. The sum of labeled and unlabeled large lymphoid cells per standard class of DNA content ( x ± 0.2 c) was greatest in a class around the 2 c mode, and smallest in classes of from 3.0 to 3.8 c DNA content. All large blasts with a relative DNA content of between 2.5 and 3.8 c were labeled indicating that throughout the major part of S phase there was no appreciable interruption in nuclear DNA synthesis. The median grain count per labeled large lymphoid blast was lowest in cells with a DNA content near the 2 c and 4 c modes, respectively, and highest in elements with a DNA content approximating 3.6 c. The inverse relative number of large lymphoid cells per class of DNA content paralleled satisfactorily the median grain count per labeled cell in the same class. These results are interpreted as indicating that in this particular cell system in vivo (1) unlabeled blasts with a DNA content near the 2 c mode are comprised of both elements in G 1 and some in early S phase; (2) nuclear size increases as cells develop from G 1 to G 2 phase although the nuclear surface area of an individual lymphoid cell in smears does not permit classification with regard to subphases of the division cycle; (3) throughout the major part of S phase overall nuclear DNA synthesis rate is rather adequately reflected by incorporation of 3H-thymidine suggesting that for the latter, possible changes during S phase of endogenous precursor pool sizes and/or activities of enzymes instrumental in incorporation of 3H-thymidine into DNA are of minor importance; (4) integrated nuclear DNA synthesis rate is highest in the second half and most probably lowest at both extremes of the S period.