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PubMed1993

Enhancement of LAK-like activity and cytokine induction in regional lymph nodes and spleen cells of mice after intralymphnodal injection of OK-432, a killed streptococcal preparation.

A single dose of inactivated streptococci (OK-432) was injected into the popliteal lymph nodes of male CDF1 mice and its effects on popliteal, inguinal, and para-aortic lymph node cells and spleen cells were investigated and compared with the effects of subcutaneous injections of the same dosage of OK-432. Regional lymph node cells and spleen cells obtained from intralymphnodally injected mice lysed not only natural killer (NK)-sensitive YAC-1 cells, but also NK-resistant P-815 and meth-A cells. Lysis of target cells was inhibited when effector cells were treated with anti-Thy-1.2 or anti-Lyt-2.2 monoclonal antibody and complement, but no inhibition was apparent after treatment with anti-asialo-GM1 or anti-Lyt-1.2 antibody and complement. These results suggest that the effector cells are lymphocyte-activated killer (LAK) cells. An enhanced capacity of lymph node cells to produce cytokines, tumor necrosis factor and interleukin 1 upon restimulation with lipopolysaccharide was found only in intralymphnodally injected mice. Thus, the induction of LAK-like cells and cytokine production in regional lymph nodes and spleen cells by the intralymphnodal administration of OK-432 should be effective for the inhibition or treatment of lymph node metastases.

Animals↗
PubMed1994

Phase Ib trial of the effect of peritumoral and intranodal injections of interleukin-2 in patients with advanced squamous cell carcinoma of the head and neck: an Eastern Cooperative Oncology Group trial.

Thirty-six patients with unresectable squamous cell carcinoma of the head and neck were entered into a phase Ib trial evaluating the toxicity, maximally tolerated dose (MTD), and immunomodulating effects of locally administered interleukin-2 (IL-2). Patients received daily IL-2 injected perilesionally in divided doses in each of four quadrants and bilaterally into the superior jugular lymph nodes. The dose of IL-2 began at 200 U/day and was escalated to 4 x 10(6) U/day in groups of six patients. Overall, regionally administered IL-2 was well tolerated. The most frequently encountered toxicities were fever, hepatotoxicity, and hypotension. Dose-limiting toxicity was encountered at 4 x 10(6) U. Of the 36 patients treated, 2 partial responses were noted at 2,000 and 4 x 10(6) U. We conclude that regionally administered IL-2 is well tolerated in patients with head and neck cancer and that the MTD is 2 x 10(6) U/day, similar to what has been reported with systemically administered IL-2. Although the overall response rate was low, it may be improved with prolonged administration of IL-2 or by combining it with other biologic or cytotoxic agents.

Adult↗
PubMedDate not supplied

Application of direct cannulation and injection lymphangiography to the study of the canine cardiac and pulmonary efferent mediastinal lymphatics.

Lymphangiograms of canine cardiac and pulmonary efferent mediastinal lymphatics were made by cannulation and injection of Ethiodol. Injections were made singly and serially. The mediastinal lymphatics and lymph nodes, which constitute the pathways of drainage of the heart and lungs, were delineated from the point of cannulation to the right and left inferior cervical region where the right lymphatic duct and thoracic duct are located. Lymphangiography reveals that the lymphatics which drain the heart and lungs may join to form common mediastinal lymphatic channels. Interconnections between mediastinal channels were demonstrated. The lymphatics terminated in the region of both the right lymphatic duct and thoracic duct in every subject. The so-called "cardiac node of Drinker" is usually a group of pretracheal nodes rather than a single node. The pretracheal nodes and those more cephalad receive drainage of lymph from both the heart and lungs. These studies suggest that lymph collected by cannulation of a "cardiac" lymphatic adjacent to the "cardiac node" will contain pulmonary as well as cardiac lymph. Thus the high flows reported by many investigators for "cardiac" lymph probably indicates that pulmonary lymph is mixed with cardiac lymph, and that the experimental data should be interpreted with this in mind.

Animals↗
PubMed1995

Intrathymic tolerance in the Lewis-to-F344 chronic cardiac allograft rejection model.

Successful induction of donor-specific unresponsiveness by intrathymic inoculation of alloantigen in several experimental acute rejection models has led us to hypothesize that similar immune manipulations can prevent chronic rejection and development of graft arteriosclerosis in the Lewis-to-F344 rat chronic cardiac allograft rejection model. Recipient F344 rats were treated with donor (Lewis) splenocytes by intrathymic injection (i.t.) alone (10 x 10(6) cells/lobe); with donor splenocytes i.t. plus a one-time dose of ALS (1 mg) by intraperitoneal injection (i.p.); or with ALS i.p. (1 mg) alone 2 and 6 weeks prior to heterotopic Lewis heart transplantation. Control F344 recipients received saline i.t. Allografts were monitored by daily palpation, and long-term surviving grafts were harvested on day 90 for histopathologic analysis. Control allografts had 28.6% long-term survival (> 90 days) with mean graft survival of 46.7 +/- 12.2 days. At day 90 the surviving control allografts were enlarged and fibrotic with barely palpable heartbeat (mean heartbeat grade 0.29 +/- 0.18), and histologically showed diffuse moderate mononuclear cell infiltrates and advanced graft arteriosclerosis (mean vessel score 3.57 +/- 0.10 and 89 +/- 1% vessels diseased). Recipient treatment with intrathymic donor splenocytes alone significantly prolonged graft survival (89% long-term survival; mean 83.8 +/- 6.2 days, P < 0.04), but did not significantly inhibit the development of graft arteriosclerosis (score 2.98 +/- 0.53 and 79 +/- 8% diseased, P = NS). By contrast, treatment with i.t. donor splenocytes plus ALS 2 weeks prior to transplantation prolonged graft survival (100% long-term; mean 90.0 +/- 0.0 days, P < 0.04), and markedly inhibited graft arteriosclerosis (score 0.80 +/- 0.14, P < 0.05; 27 +/- 4% diseased, P < 0.05). ALS alone given two weeks prior to transplantation also prolonged graft survival (100% long-term; mean 90.0 +/- 0.0 days, P < 0.04), and inhibited graft arteriosclerosis (score 0.89 +/- 0.31, P < 0.05; 25 +/- 7% diseased, P < 0.05). However, when ALS was given 6 weeks prior to heart transplantation the beneficial effect of ALS alone was abolished, suggesting that lymphocyte depletion may have been responsible for the observed effects when ALS was administered at 2 weeks. Interestingly, intrathymic donor splenocytes plus ALS 6 weeks prior to transplantation, on the other hand, showed significant prolongation of allograft survival (100% long-term, mean 90.0 +/- 0.0 days, P < 0.04), and inhibited graft arteriosclerosis (score 0.41 +/- 0.02, P < 0.05; 16 +/- 2% diseased, P < 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1996

Acquired systemic tolerance to rat cardiac allografts induced by intrathymic inoculation of synthetic polymorphic MHC class I allopeptides.

This study extends the finding that intrathymic (IT) injection of 3M KC1 extracts of T cells induces transplant tolerance to the use of well defined polymorphic MHC class I allopeptides derived from the hypervariable domain of RT1.Au (WF MHC class I). While three of the six synthetic RT1.Au peptides were immunogenic, three others were nonimmunogenic when tested in ACI responders. In our initial studies, we examined the effects of IT injection of a mixture of equal concentrations of the three nonimmunogenic RT1.Au peptides on WF cardiac allograft survival in ACI recipients. The results showed that a single IT injection of 100 and 300 microg class I MHC allopeptides on day -7 relative to cardiac transplant did not significantly prolong graft survival in naive ACI recipients (MST of 9.8, and 12.3 days vs. 10.5 days in controls). In contrast, 600 microg allopeptides injected IT resulted in modest prolongation of graft to an MST of 19.5 days. However, IT injection of 600 microg allopeptides combined with 0.5 ml ALS on day -7 led to permanent acceptance (>200 days) of cardiac allografts in 7/9 ACI recipients compared with survival of 24.2 days in ALS alone treated controls. In contrast, similar treatment led to acute rejection of third party (Lewis) cardiac allografts. Intravenous injection of 600 microg allopeptides combined with ALS did not result in prolonged graft survival (26.8 days). The long-term unresponsive ACI recipients (>100 days) challenged with second-set cardiac grafts accepted permanently donor-type (WF) grafts while rejecting the third party (Lewis) grafts, a finding that confirms acquired systemic tolerance. These findings confirm the role of IT injection of synthetic polymorphic allopeptides in the induction of acquired thymic tolerance and provide the rationale for testing this strategy in large animals and eventually in man.

Amino Acid Sequence↗
PubMed1997

Fat absorption after small intestinal transplantation in the rat.

BACKGROUND: Intestinal transplantation is now used for patients with severe malabsorption, however, little data exists quantifying the ability of the graft to absorb fat. This study tested the hypothesis that intestinal transplantation would not affect the lymphatic or venous uptake of fatty acids. METHODS: A syngeneic rat model of intestinal transplantation (SIT) with caval drainage of the graft was used. Control animals underwent intestinal division and reanastomosis (n=15 in each group). The animals were followed for 6 weeks, and fat absorption in vivo was quantified. The animals were anesthetized, sampling catheters were placed in the jugular and superior mesenteric veins and in the mesenteric lymphatic duct, and a feeding tube was passed into the duodenum. Animals were allowed to recover, and a steady-state duodenal infusion of lauric (C12:0) and palmitic (C16:0) fatty acid emulsion was begun. A radiolabeled pulse of lauric (C12:0) and palmitic (C16:0) fatty acid was then given, and the subsequent appearance in the lymphatic and venous systems was quantified. RESULTS: In vivo absorption of dietary fat was preserved, but after transplantation the mesenteric lymphatic flow and cumulative lymphatic appearance of both labels was significantly reduced (flow reduced from 4.8+/-1.1 in controls to 1.0+/-0.29 ml/hr in transplant animals, whereas lauric acid absorption was 33+/-11.4% in controls vs. 7.5+/-2.5% in transplant animals). There was a modest increase in the jugular venous appearance of the fatty acids (2.0+/-1.1% in transplant animals vs. 0.75+/-0.55% in controls for lauric acid; P<0.05 for all comparisons). Absorption of lauric and palmitic acids was very similar, and there was no preferential absorption detected in the portal venous system. Dye studies demonstrated lymphatic recannulization around the vascular anastomosis, into the retroperitoneum. CONCLUSIONS: These results suggest that in this model of SIT, fat absorption via the mesenteric duct is reduced, but that compensatory collaterals form into the retroperitoneal lymphatics. There was no evidence of any significant increase in portal venous uptake of fatty acids after SIT, nor of preferential absorption of medium-chain fatty acids. These results may have implications for patients after SIT.

Animals↗
PubMed2004

Intralabyrinthine fluid dynamics: Meniere disease.

PURPOSE OF REVIEW: Meniere disease has long been postulated to be a disorder of intralabyrinthine fluid dynamics. RECENT FINDINGS: More recent developments in this field indicate that the control of fluid movement may be at a cellular level and that hormonal influence may be important. SUMMARY: The control of fluid and ion movements through aquaporins and gap junctions in the cell membranes are creating new perspectives in the mechanism of development of endolymphatic hydrops as well as potential methods for treatment. Intralabyrinthine fluid dynamics also play a role in the ability to locally deliver drugs to the inner ear through the middle ear.

Combined Modality Therapy↗
PubMed1997

Intranodal injection of anticancer drugs into fixed cervical metastatic lymph nodes.

OBJECTIVE: In patients with head and neck carcinoma, fixed enlarged metastatic lymph nodes (LNs) are sometimes inoperable and carry an increased risk of mortality. To control metastatic LNs, we attempted intranodal injection of anticancer agents. SUBJECTS AND METHODS: Fifteen patients with squamous cell carcinoma arising in the gingiva (8), tongue (3), floor of the mouth (1), or maxillary sinus (3) were enrolled. These patients consisted of two groups, those in the early era in which the fixed LNs of six patients were treated with 60Co (RA group) and those in the late era in which both radiation and intranodal injection of anticancer agents were administered to nine patients (IN group). Intranodal injection consisted of peplomycin, 5-fluorouracil, and cis-diamminedichloroplatinum. RESULTS: In the IN group, LNs regressed from about 40% to nearly 100%, although two patients showed no appreciable response. The LNs treated by combination therapy regressed considerably while LNs in the same patients treated with 60Co alone showed a minor response or grew gradually. In three patients, the LNs regressed sufficiently to be extirpated safely. The good clinical response in the locally injected LNs was histologically associated with distinct evidence of tumor cell degeneration. In the RA group, none of the LNs responded to radiation with 60Co; one LN exhibited slight regression, but the others enlarged during and soon after the radiation. Compatible with the clinical effects, many patients in the IN group demonstrated a good prognosis; three are alive without disease, and four survived for prolonged periods. However, all patients in the RA group died due to progression of the positive LNs or pulmonary complication within 10 months. CONCLUSION: These results indicate that intranodal injection of anticancer drugs is useful for the management of fixed enlarged LNs.

Aged↗
PubMed1991

Histological findings in rabbit lymph nodes after endolymphatic injection of liposomes containing blue dye.

Liposomes produced from phosphatidylcholine and cholesterol and containing Patent Blue V have been injected endolymphatically in the rabbit. The lymph nodes stained dark blue and retained this colour until termination of the experiment after 28 days. No toxic effects were observed histologically. The liposomes were deposited within the macrophages in a fine granular pattern. Because endolymphatic injection of such liposomes produces excellent contrast between retroperitoneal lymph nodes and their surrounding fatty tissue, it can improve both the radicality and the selectivity of lymphonodectomy in oncological surgery.

Animals↗
PubMed2006

Replication and propagation of attenuated vesicular stomatitis virus vectors in vivo: vector spread correlates with induction of immune responses and persistence of genomic RNA.

Live-attenuated vesicular stomatitis virus (VSV) vectors expressing foreign antigens induce potent immune responses and protect against viral diseases in animal models. Highly attenuated (VSV-CT1) or single-cycle VSV (VSVDeltaG) vectors induce immune responses lower than those generated by attenuated wild-type VSV vectors when given intranasally. We show here that reduced spread of the more highly attenuated or single-cycle vectors to other organs, including lymph nodes, correlates with the reduction in the immune responses. A reverse transcription, real-time PCR assay for VSV genomic RNA (gRNA) sequences showed long-term persistence of gRNA from replicating vectors in lymph nodes, long after viral clearance. Such persistence may be important for induction of potent immune responses by VSV vectors.

Animals↗
PubMed2000

Local application of capsaicin into the draining lymph nodes attenuates expression of adjuvant-induced arthritis.

Adjuvant-induced experimental arthritis (AA) was examined in adult male Lewis rats after isolated capsaicin (CAPS)-induced loss of small, nonmyelinated, afferent fibers in lymph nodes draining the site of adjuvant challenge. AA was induced by intradermal injection of Freund's complete adjuvant (CFA) into the subplantar area of the right hind paw. Controls received similar injections of mineral oil, the vehicle for CFA. One day later, half of the CFA-treated rats and half of the mineral oil-treated rats received injections of CAPS bilaterally into the draining lymph nodes (DLN). The DLN of remaining rats were injected with 50:50 ethanol/sterile physiological saline, the vehicle for CAPS. This paradigm resulted in four groups designated: CFA/CAPS, CFA/vehicle, vehicle/CAPS and vehicle/vehicle. Since substance P (SP) is present in small, nonmyelinated, afferent fibers, the target of the neurotoxin, CAPS, a radioimmunoassay specific for SP was used to verify the loss of these nerve fibers. CAPS injections into the DLN resulted in a loss in SP concentration in the DLN, with no depletion of SP in the spleen or sciatic nerve. These findings support the destruction of SP-containing nerves, which we interpret as verification of the selective loss of small, non-myelinated afferent nerves in the DLN with no significant spread of the neurotoxin to the nearby sciatic nerves which supply small, nonmyelinated, afferent fibers to the hind limb joints. Also, preservation of SP content in spleen indicates CAPS did not circulate via the lymphatic drainage. No chronic inflammation was observed in the fore or hind limbs from rats treated with the vehicle for CFA (vehicle/vehicle, vehicle/CAPS) at any time during the study. In CFA/vehicle-treated rats, bilateral, symmetrical inflammation of the hind limbs was apparent 14 days after challenge with CFA, and became progressively more inflamed through day 20. In contrast, hind limb inflammation in arthritic rats treated with CAPS was not symmetrical. On days 14 and 20 after challenge with CFA, the inflammatory response in the left hind limb, contralateral to the site of CFA injection, was significantly (p < 0.05) attenuated compared with the response seen on the right side of CFA/CAPS-treated rats, and with the response seen in left hind limb of CFA/vehicle-treated animals. In fact, the mean dorsoplantar width of contralateral hind limbs from CFA/CAPS-treated animals was not different from that measured in non-AA control groups. These findings support a role for small, nonmyelinated, sensory nerves that modulate immune responses in DLN in the development and progression of AA in Lewis rats.

Afferent Pathways↗
PubMed1976

Effect of glucose administration on the blood sugar and pancreatic islets of the frog, Rana tigrina.

Glucose evoked significant hyperglycemia in the frog, Rana tigrina, 0.5 h after the injection and it took a longer time to regain normoglycemia than mammals, thus exhibiting a low glucose tolerance. The islet tissue of injected animals showed degranulation and atrophy of beta-cells only, the alpha-cells remained almost normal. This damage suggests that the beta-cells of this frog are functionally like those of mammals and other vertebrates, and secrete insulin. A single dose of glucose was found incapable of producing permanent diabetes, and beta-cells damage was repairable at later stages. Excess of glucose in the blood seems to stimulate secretion of insulin by the beta-cells.

Animals↗
PubMed1997

Distribution and ultrastructure of the stomata connecting the pleural cavity with lymphatics in the rat costal pleura.

We investigated the detailed distribution and ultrastructure of the stomata connecting the pleural cavity and the lymphatics in the rat costal pleura by scanning electron, transmission electron and light microscopy. The mesothelial cells lining the costal pleura appeared as both flattened and thick cell bodies. The thick cells possessed more rough endoplasmic reticula, Golgi complexes, mitochondria, and free ribosomes than the flattened cells. The thick cells were distributed in the intercostal regions each cephalic to the junction of the costal cartilage and bone, and in the band-like regions along the cephalic and caudal sides of each rib in the lateral and dorsal thoracic walls. In the regions lined with thick cells, there were stomata [12.9 +/- 10.3 microns2 (mean +/- SD) in area] consisting of prolongations of thick mesothelial cells and funnel-like projections of lymphatic endothelial cells that came up along the rims of the pores (5.9 +/- 3.2 microns2 in average area) in the submesothelial collagen fiber network. At the stomata, the basal lamina of the mesothelium was continuous with that of the endothelium. The mesothelial cells forming the stomata were mostly in close contact with the endothelial cells, but some gaps also existed between them. Valve-like endothelial flaps were frequently observed wherever endothelial cells constituting the stomata merged into the submesothelial lymphatics. Also present were lymphatic bulges that were either in close contact with the base of the thick mesothelial cells or exposed through the mesothelial pores. The lymphatic network was especially well developed in the submesothelial layer at and around the thick-cell regions. The initial lymphatics drained into the intercostal collecting lymphatics, which in turn led into either the parasternal or paravertebral lymphatic trunk. Our results suggest that the stomata play a major role in absorbing fluids and particulates in the pleural cavity. The thick mesothelial cells appear to secrete chemotactic substances to the endothelial cells. Understanding the heterogeneous distribution of the stomata could prove to be important clinically in inflammatory diseases and tumors in the chest.

Animals↗
PubMedDate not supplied

Flow velocity of cutaneous lymphatic capillaries in patients with primary lymphedema.

For the first time measurements of lymph flow velocities in cutaneous microlymphatics of patients with lymphedema were performed and compared with healthy subjects. Flow velocity in single lymphatic skin capillaries was measured using fluorescence video microscopy after subepidermal microinjection of FITC-dextran 150,000 in 15 healthy volunteers and 16 patients with primary lymphedema. Initial filling of the lymphatic capillary network was fast with significantly higher mean velocities in patients with primary lymphedema than in healthy controls (890 +/- 43 vs. 550 +/- 390 microns/s, p < 0.05). The resting velocities were not significantly different between controls and patients (10.3 +/- 4.1 vs. 16.6 +/- 13.9 microns). In 12 out of the 16 lymphedema patients cutaneous backflow of the fluorescent contrast medium from deeper invisible lymphatics was observed. In 4 of these patients rhythmic reflux with a mean frequency of 1.4 +/- 0.5 cycles/min was measured by video densitometry in microlymphatics with a significantly (p < 0.01) enhanced diameter. Mean flow velocity (Vp) in these precollectors was significantly increased compared to the resting velocities (p < 0.01). On the basis of these results the hypothesis is advanced that rhythmic cutaneous backflow originates from intrinsic contractions of deeper lymph collector segments and is transmitted to the superficial microlymphatics through incompetent connecting channels. This newly recognized mechanism appears to be an important factor for the pathophysiology of lymphedema.

Adult↗
PubMed1988

Carcinogenicity testing of black pepper (Piper nigrum) using the Egyptian toad (Bufo regularis) as a quick biological test animal.

Milled black pepper (Piper nigrum) force-fed to Egyptian toads as a suspension in amphibian saline or injected subcutaneously in the dorsal lymph sac as an ethanol extract, induced primary tumours in the liver and secondary tumours in other organs (kidney and spleen). When applied to the skin of experimental animals as an ethanol extract, black pepper induced primary tumours in the liver and secondary tumours in the ileum and stomach. Tumours of the liver were diagnosed as hepatocellular carcinomas and those of the other organs as metastases of the primary liver tumours. It is speculated that one or more constituents of black pepper may be responsible for tumour induction in the organs of the Egyptian toad Bufo regularis.

Administration, Cutaneous↗
PubMed1976

Reevaluation of inguinal lymph node injection for production of adjuvant arthritis in the rat.

An experiment was designed to compare the efficiency of lymph node injection for the induction of adjuvant arthritis (AA) with that of conventional footpad injection in the rat. Quantitative studies revealed that the minimal dose required for induction of AA by the lymph node route is one fifth of that by the footpad route. Thus, the lymph node route was found to be more efficient than the footpad method in terms of higher incidence and earlier onset of AA. PPD in Freund's incomplete adjuvant was able to produce tuberculin sensitization in the rat. The lymph node route again proved to be superior in terms of consistent appearance of the 24-hour reaction on days 8 and 14 and prolongation of the skin reaction over 48 h. These findings show that the lymph node method is so efficient in the rat that it will be especially useful for the trial induction of AA with various materials of unknown potency as well as for production of delayed hypersensitivity. In addition, this injection method appears to be a simple and efficient technique for assay of other immunological reactions.

Animals↗