Morphologic alterations in canine marrow of long-term survivors after 1200 R whole-body x-irradiation and autologous blood leukocyte engraftment.

The marrow matrix of total-body x-irradiated dogs (1200 R midline dose) was able to support effective hemopoiesis for several hundred days if the animals were transfused with their own mononuclear leukocytes collected from the blood prior to irradiation and preserved at ultralow temperatures. However, a lesion developed in the marrow, consisting of a fibrosis originating in conjunction with or from the endosteum. The fibrotic tissue substantially reduced the available marrow space in dogs with advanced lesions. The number of autologous, cryopreserved mononuclear leukocytes transfused ranged from 0.32 X 10(9) to 1.63 X 10(9)/kg body weight. The observation period extended to a maximum of 898 days after irradiation.     

Systemic suppression of human peripheral blood phagocytic leukocytes after whole-body UVB irradiation.

We examined systemic effects of whole-body UVB irradiation on human peripheral blood phagocytes. We found that 24 h after a single erythemal dose of UVB radiation two phagocyte functions, adhesion and phagocytosis, were reduced by 50%. This functional suppression was accompanied by a significant decrease in the expression of complement receptors (CR1 and CR3) and IgG Fc receptors (FcRII and FcRIII). The greatest reduction (47%) was observed in CR3, which is important for both adhesion and phagocytosis. A kinetic analysis showed that both CR1 and CR3 levels started to decrease 15 min after the UVB exposure, reaching the lowest levels at 4.5- and 24-h time points, respectively. The down-modulation of CRs after whole-body UVB exposure was not due to a defective receptor synthesis or translocation from internal stores to plasma membrane because the maximal CR levels in stimulated cells were not affected by UVB. No change in the serum soluble ICAM-1 was detected after UVB, which rules out CD1 1b epitope masking by sICAM-1. UVB did not release low-receptor-density myeloid progenitor cells from storage pools into circulation. Interleukin 10, a mediator of UVB-induced immunosuppression, was unable to modulate CR expression in vitro. When seven suberythemal whole-body UVB exposures were given repeatedly within 2 weeks, a significant decrease in CR expression was seen, which was greatest after three irradiations. Our data suggest that an exposure to UVB has systemic effects in humans which, possibly due to the down-modulation of preexisting cell-surface receptors, suppress some important functions of circulating phagocytic cells.     

The mononuclear cells of human mesenteric blood, intestinal mucosa and mesenteric lymph nodes: compartmentalization of NK cells.

The proportions of T cell subsets and Leu 7+ cells and the spontaneous cell-mediated cytotoxicity (SCMC) of isolated mononuclear cells have been determined across the mesenteric vascular bed and along the intestinal mucosal-mesenteric lymph node (MLN) axis in patients undergoing abdominal surgery. Whereas the proportion of T4+ and T8+ cells were similar in simultaneously taken PVB and mesenteric venous blood (MVB), the proportion of Leu 7+ cells was higher in MVB in 16 of 17 studies (15.4 +/- 6.8%, 10.8 +/- 5.1%). Additional studies showed that the proportions of lymphocyte subsets in peripheral arterial blood are the same as those in PVB. Thus, an enrichment of Leu 7+ cells occurs across the mesenteric vascular bed. Isolated intestinal and MLN mononuclear cells contained similarly high proportions of T4+ and T8+ cells as in PVB but Leu 7+ cells made up a minority subpopulation in intestinal (1.3 +/- 0.8%) and MLN mononuclear cells (1.0 +/- 0.9%). The SCMC of intestinal and MLN mononuclear cells was low and paralleled the proportion of Leu 7+ cells. Despite the higher proportions of Leu 7+ cells in MVB, the SCMC was less than that of PVB in eight patients with inflamed intestine and not significantly different from PVB in seven patients with normal intestines. These paradoxical findings were at least in part due to inhibitory factors in mesenteric plasma. In conclusion, NK cells appear to be largely confined within the vascular system and the enrichment of Leu 7+ cells across the mesenteric vascular bed suggests that this compartmentalization may be due to differences in the traffic of lymphocyte subpopulations through the intestinal mucosa and MLN.     

Functional anatomy of lymph nodes. II. Peripheral lymph-borne mononuclear cells

In the rabbit a number of large mononuclear cells with ruffled surface membranes travel from the skin and superficial tissues of the leg, via the lymphatics, to the popliteal lymph node: they constitute 40-50% of the total cell population in the afferent lymph. About 10% of these cells are actively phagocytic when tested in vitro and about 3% are found to contain Langerhans granules. After isotopic labelling the majority of lymph-borne mononuclear cells can be detected within the regional node for at least 24 hours; most being located in the paracortex and a few in the interfollicular cortex. It is proposed that these cells, including those containing Langerhans granules, belong to the "mononuclear phagocyte system." Possible functions of these lymph-borne cells are discussed with particular reference to antigen transport.     

Modification of lymph by lymph nodes. II. Effect of increased lymph node venous blood pressure

A previous study from this laboratory demonstrated that lymph nodes can change the protein concentration and colloid osmotic pressure of lymph by transfer of protein-free fluid between the blood and lymph compartments. In that study a Starling force disequilibrium across the blood-lymph barrier caused fluid to transfer through the barrier in the direction required to establish equilibrium of Starling forces. In the present study we examined the effect of increased lymph node venous blood pressure on efferent lymph protein concentration and efferent lymph flow. We utilized an isolated dog popliteal lymph node preparation in which afferent lymph having various protein concentrations was perfused into the node at an average flow rate of 19.1 +/- 0.3 (SD) microliter/min. We compared steady-state values of prenodal and postnodal lymph flows and protein concentrations during various steady-state levels of lymph node venous blood pressure. When venous pressure was increased, the protein concentration of the efferent lymph invariably decreased and the efferent flow rate invariably increased. Measurements showed that an average of 96% of the change in lymph protein concentration was caused by transfer of protein-free fluid through the lymph node blood-lymph barrier. The results of this study indicate again that the lymph node functions as a fluid exchange chamber in which fluid is transferred between the blood and lymph compartments in the direction required to establish equilibrium of the Starling forces across the blood-lymph barrier.     

Observations of the early diuretic response after intravenous administration of bumetanide and furosemide in dogs.

A transient natriuretic peak was observed in dogs the third minute after the i.v. administration of the highly active diuretics bumetanide and furosemide. The peak is dependent on the sodium balance such that it is potentiated during positive sodium balance and is not observed in sodium depleted dogs. A relationship exists between the simultaneous occurrence of the peak and abolition of the cortico-medullary electrolyte gradient.     

Changes in DNA and surface properties of peripheral blood leukocytes in monkeys after whole-body γ-irradiation

Central Roentgeno-Radiologic Research Institute, Ministry of Health of the USSR, Leningrad. Institute of Biophysics, Ministry of Health of the USSR, Moscow. Research Institute of Experimental Pathology and Therapy, Academy of Medical Sciences of the USSR, Sukhumi. (Presented by Academician of the Academy of Medical Sciences of the USSR V. A. Lapin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 108, No. 8, pp. 158–159, August, 1989.     

Quantitative evaluation of Epstein-Barr-virus-infected mononuclear peripheral blood leukocytes in infectious mononucleosis.

We devised a quantitative assay for Epstein-Barr-virus-infected mononuclear leukocytes (virocytes) to determine their prevalence in the blood of patients with acute-phase and convalescent-phase infectious mononucleosis and in healthy Epstein-Barr-virus-seropositive controls. Mononuclear peripheral blood leukocyte suspensions were tested for virus-determined cytoproliferative activity by cocultivation with human cord-cell indicator cultures. The highest levels of virocytes among circulating mononuclear leukocytes were found in the early acute phase of infectious mononucleosis (up to 0.05 per cent). Virocytemia decreased to levels comparable with those of healthy controls (less than 0.00001 per cent) by the third month after onset of infectious mononucleosis. These findings provide a quantitative profile of the course of the infection at cellular level and support existing evidence of the efficiency of immune control mechanisms in limiting Epstein-Barr-virus infection during the course of infectious mononucleosis.     

Detection of human cytomegalovirus antigen and DNA in lymph nodes and peripheral blood mononuclear cells of patients with angioimmunoblastic lymphadenopathy with dysproteinemia.

The cause of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) remains unknown. It is characterized by acute onset, severe constitutional symptoms, cervical or generalized lymphadenopathy, lymphopenia, and polyclonal hypergammaglobulinemia, all of which are highly suggestive of a viral origin. Using immunohistochemical methods, employing murine monoclonal antibody as the primary antibody, we detected human cytomegalovirus antigen in the lymph nodes of eight of 11 patients with AILD. Cytomegalovirus DNA was also detected in the peripheral blood mononuclear cells by DNA dot hybridization in all five of the patients with AILD who were tested using this technique. None of the lymph nodes from the 11 patients stained positive for the rubella virus antigen. Based on the above evidence and the similarity of the immunologic abnormalities found in both AILD and cytomegalovirus infection, the possible role of cytomegalovirus as one of the causative agents for AILD is proposed.     

Electron-microscopic study of plasma cells from regional lymph nodes of rats after topical interferon administration

Transmission electron microscopy of plasma cells from soft strands of cranial deep cervical lymph nodes removed from male Wistar rats at different times after their intranasal treatment with an aqueous α-interferon solution showed a decreased volume of the Golgi complex and lowered numbers of ribosomes and mitochondria in both mature and immature plasma cells. The results of the study are tentatively interpreted as indicating reduced energy potentials and antigen-synthesizing capabilities of the plasma cells formedde novo after α-interferon treatment. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 7, pp. 60–63, July, 1996     

The sequence of changes in blood flow and lymphocyte influx to stimulated rat lymph nodes.

The rat popliteal lymph node was studied from 1 hr to 8 days after the footpad injection of either sheep erythrocytes or syngeneic rat erythrocytes. The following were measured relative to the contralateral (unstimulated) lymph node: (i) blood flow; (ii) lymph node weight; (iii) influx of lymphocytes from the blood; (iv) [3H]-thymidine incorporation; (v) [35S]-sulphate incorporation into macromolecular form (chiefly by high endothelial venules). After the arrival of sheep erythrocytes all five quantities showed substantial increases which began in a definite sequence. The blood flow started to rise first and may have been the main factor contributing to the later increase in lymphocyte influx. Increased sulphate incorporation began later than the rise in lymphocyte influx. After the injection of rat erythrocytes a small increase in lymphocyte influx was found without a corresponding increase in blood flow. In rats irradiated before the footpad injections lymphocyte influx increased three-fold after sheep erythrocytes, rat erythrocytes or PBS, again without a corresponding increase in blood flow. Thus while variation in blood flow to high endothelial venules is one important factor in determining the supply of lymphocytes to the lymph node other factors are operative in certain situations.     

Splenic lipidosis after administration of intravenous fat emulsions.

Spleens showing fatty infiltration and necrosis of the pulp were found at necropsy on several patients who had received intravenous fat emulsions during their terminal illnesses. The postmortem findings are described and the clinicopathological correlation is discussed with special reference to the phenomenon of creaming of the emulsion.     

The regeneration of lymph nodes after their partial removal or injury

Summary A series of experiments were conducted on albino rats, weighing 100–130 g It was established that fliac lymphatic nodes are able to regenerate after injury or removal of their greater part. However, regeneration takes place only in conditions of continued circulation of lymph in the node or in relatively rapid recovery of the circulation. In stable obstruction of lymph drainage, on the contrary, there is no regeneration in the lymph node and gradual fibrosis takes place in its residual part.Presented by Academician A. D. Speransky     

Lymphocyte emigration from lymph nodes by blood in the pig and efferent lymph in the sheep.

Two types of experiment using local labeling of lymph nodes with FITC showed that lymphocytes emigrate from lymph nodes, predominantly in blood in the pig and in efferent lymph in the sheep. In the first type of experiment with the pig, few cells emigrated via the lymph, while the number of labelled cells in the blood increased progressively and the indices in mesenteric blood were always higher than in jugular blood in simultaneously-drawn samples. However, in the sheep, when efferent lymph flowed freely, very low numbers emerged in blood and continuing large numbers of lymphocytes emerged in efferent lymph. In the second type of experiment carried out wholely under anaesthetic on mesenteric lymph nodes in pigs and sheep, and on superficial inguinal lymph nodes in pigs, the lymph node was isolated, the lymph and venous drainage collected and only the arterial supply maintained. Large numbers of FITC+ lymphocytes emigrated via the vein in pigs with either node cannulation (i.e. up to 7% blood lymphocytes were labelled with an emigration rate of approximately 10(8) cells/hr) but in sheep, while lymph contained approximately 30-80% labelled cells and the emigration rate was also approximately 10(8) cells/hr, the mesenteric blood contained very few labelled cells (approximately 0.2%, giving a mean venous emigration rate of 2.7 X 10(6)/hr). Study of the type of lymphocytes emerging from labelled pig lymph nodes and spleen during the phase of major emigration showed that sIg+ B and E rosette-forming T cells, but almost no Null cells, are involved.     

Evidence of antibody production in the rat cervical lymph nodes after antigen administration into the cerebrospinal fluid

We previously showed histologically that, in the rat, the cerebrospinal fluid drains from the subarachnoid space along the olfactory nerves to the nasal lymphatics and empties into the superficial and deep cervical lymph nodes. The present study was performed to investigate whether these lymph nodes play a role in the immune response of the central nervous system. For this purpose, keyhole limpet hemocyanin conjugated with fluorescein isothiocyanate (KLH-FITC) was administered into the subarachnoid space of the rat brain, and the time-kinetics and location of FITC and anti-FITC antibody forming cells in the cervical lymph nodes were studied histologically and immunohistochemically. FITC fluorescence was detected in superficial and deep cervical lymph nodes as well as the subarachnoid space and the nasal mucosa 2 h after FITC-KLH injection into the subarachnoid space. The specific antibody-forming cells first appeared in both the superficial and deep cervical lymph nodes on the 4th day after antigen administration although the reaction was more intense in the deep than in the superficial cervical lymph nodes. These cells were located in the medullary cords of the cervical lymph nodes. The number of antibody forming cells increased thereafter, reached a peak around the day 6, and then declined on day 10. These findings indicate that antigens introduced in the cerebrospinal fluid are drained into the cervical lymph nodes through the nasal lymphatics and initiate the antigen-specific immune response there. Thus, the cervical lymph nodes probably act as a monitoring site for cerebrospinal fluid and play a major role in the central nervous system immune response.     

Glucose and lactate kinetics after endotoxin administration in dogs.

We studied the effects of E. coli endotoxin on the glucose and lactate kinetics in dogs by means of the primed constant infusion of [6(-3)H] glucose and Na-L-(+)-[U-14C] lactate. The infusion of endotoxin induced a transient hyperglycemic level, followed by a steady fall in plasma glucose to hypoglycemic levels. The rate of appearance (Ra) and the rate of disappearance (Rd) of glucose were both significantly elevated (P less than .05) for 150 min after endotoxin, after which neither differed from the preinfusion value. The metabolic clearance rate of glucose was significantly elevated at all times 30 min postendotoxin. By 30 min postendotoxin, Ra and Rd of lactate, plasma lactate concentration, and the percent of glucose turnover originating from lactate were significantly elevated and remained so for the duration of the experiment. We concluded that after endotoxin hypoglycemia developed because of an enhanced peripheral uptake of glucose and a failure of the liver to maintain an increased Ra of glucose. We also concluded that lactate became an important precursor for gluconeogenesis and an important metabolic substrate.