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Study of DNA metabolism of lymph-node cells by direct lymphatic administration of tritiated thymidine.

A method for studying DNA metabolism in lymph-node cells by injecting tritiated thymidine intralymphatically is described. The administration of [3H]thymidine through a lymph vessel enabled a high concentration to be attained with only a small quantity of the precursor in close proximity to the cells. The significance of the method is that it may also be used in studies of metabolic processes in human lymph-nodes.

Animals↗

Interleukin-2 increases the antibody response in patients receiving autologous intralymphatic tumor cell vaccine immunotherapy.

The production of tumor-binding antibodies was studied in a group of cancer patients undergoing active specific immunotherapy with irradiated, cholesterol-treated, cell culture-derived autologous tumor cells injected by the intralymphatic route. Fifteen patients were analyzed: nine patients (four melanoma, one breast, one sarcoma, one colon, and one undifferentiated cancer) received three injections of 10 to 15 x 10(6) tumor cells, spaced 2 weeks apart, and six patients (two melanoma, two renal, one breast, and one colon cancer) received tumor cells admixed with 3 x 10(6) U recombinant interleukin-2 (IL-2) (Proleukin, Cetus, Emeryville, CA, USA) plus a 10-day intravenous infusion of 15 x 10(6) U/kg/day IL-2 after each immunization. Serum antibody binding to autologous tumor cells was measured at 2 and 4 weeks after initiation of therapy using an enzyme-linked immunosorbent assay with patient serum being added to adherent tumor cells bound to 96-well microtiter plates. After 4 weeks, we found a significant difference (0.02 less than P less than 0.04) in serum titer in the group receiving IL-2 (33% mean increase) compared with the non-IL-2 group (8% mean increase). Although neither group showed clinical improvement in response to the therapy, the results clearly demonstrated the efficacy of IL-2 in augmenting patient antibody response to autologous intralymphatic tumor cell immunization.

Antibodies, Neoplasm↗

The pathomechanism of posttraumatic edema of lower limbs: I. The effect of extravasated blood, bone marrow cells, and bacterial colonization on tissues, lymphatics, and lymph nodes.

BACKGROUND: The mechanical injury of soft tissues and bones of lower extremities is frequently followed by long-lasting edema at the site of trauma and distally. The pathomechanism of this complication remains unclear. Venous thrombosis and interruption of lymphatics are considered to be the main etiologic factors. We propose a concept that protracted healing of injured tissues and bones with involvement of the regional lymphatic system (lymphatics and nodes) is responsible for persistence of edema. The events affecting the first (scavenging) phase of healing of traumatized tissues, such as hematoma, translocation of bone marrow cells to soft tissues, and colonization by microorganisms, and in particular their effects on lymphatics and lymph nodes, were studied. METHODS: Mongrel dogs weighing 15-20 kg were used. Fresh blood or bone marrow cell (BMC) suspension was injected subcutaneously or intralymphatically into the paw. Strains of saprophyte bacteria residing on the skin surface were cultured and injected intradermally. Oil-contrast lymphography was performed before and after injections to evaluate the changes in lymphatics and nodes. Biopsy samples of paw skin, subcutaneous tissue, and regional lymph nodes (LN) were taken. The responsiveness of LN lymphocytes was studied in autologous mixed cultures with peripheral blood lymphocytes (PBL), BMC, and cultured bacteria. RESULTS: The PBL from subcutaneously injected blood were evacuated by the lymphatic route at a rate of 1-3%/6 hr. There was no thrombosis of lymphatic vessels or obstruction of LN sinusoids. The BMC evoked major inflammatory changes in both the skin and the LN. Bacteria caused local inflammation, dilatation of lymphatics, and destruction of node parenchyma. Autologous BMC and PBL stimulated LN lymphocytes in a 6-day culture. The responsiveness of lymph node lymphocytes to previously subcutaneously injected bacterial antigens was increased. CONCLUSION: The extravasated blood did not produce changes in skin, subcutaneous tissue, and lymphatics; however, it stimulated LN lymphocytes. The BMC and saprophyte bacteria caused major local and lymph node inflammatory response. All these factors may contribute to the local edema in the initial phases of healing of traumatized tissues.

Animals↗

Albumin shifts across the extracellular space secondary to experimental infections.

The pathway across tissue spaces of intravenously injected 125I albumin was studied in five dogs before and after the injection of Escherichia coli endotoxin by the use of perforated plastic capsules placed in the subcutaneous tissue. The already negative extracellular space pressure became less so after the endotoxin injection, when albumin was detected shifting from the intravascular space into the extracellular space compartment and then into the intralymphatic space. The injection of endotoxin produced a marked increase in the thoracic duct lymph flow, while, at the same time, erythrocytes entered the lymphatic stream. Results of this study suggest that experimental canine endotoxemia is associated with an increased passage of albumin into the extravascular compartment and explains, in part, the fall in serum levels of this protein during clinical and experimental sepsis.

Animals↗

The intranodal distribution of lymph-borne particles injected intravenously.

The sequential distribution of lymph-borne, i.v. injected particles of tantalum in hepatic hilar lymph nodes was studied in rats in an attempt to determine which structural compartments of a node are responsible for mechanical filtration. The injected particles reached these nodes via liver lymph but the i.v. route of administration eliminated any possibility of disturbing either lymph flow or pressure. Particles began to enter hepatic hilar nodes only after an interval of 7-8 h. They were subsequently redistributed from marginal, trabecular and medullary sinuses to the paracortex and finally to medullary cords. Particles accumulated predominantly in the paracortex at 12-24 h and thereafter in medullary cords. This sequential pattern of distribution differed significantly from that observed previously in other lymph nodes after either intralymphatic or intratissue injection.

Animals↗

Granulomatous lymphangitis. A complication of intralymphatic immunotherapy with methanol extraction residue of BCG (MER).

Patients with thick primary melanomas, (Stage I) or regional nodal spread (Stage II) are at substantial risk for recurrence following usual definitive surgical excision of the primary tumor with or without lymphadenectomy. Trials of nonspecific adjuvant agents such as bacille Calmette Guerin (BCG) in experimental animals and man suggest that antitumor effects are greatest when the agent is injected near tumor of limited mass. We report a new approach to adjuvant therapy using preoperative intralymphatic injections and intraoperative local instillations of the nonviable methanol extraction residue of bacille Calmette Guerin (MER). Thirteen individuals with thick primaries, regional metastases, or recurrent melanoma of the extremities have entered the experimental program. We report here one complication of this immunotherapy observed in four of 13 treated individuals, granulomatous lymphangitis. The clinical presentation, course, and treatment of this complication are described. Its potential relation to the success of this therapy is discussed.

Adult↗

Novel antigenic markers of human tumor regression.

The development of tumor-specific antibodies was studied in a group of cancer patients undergoing active specific immunotherapy with irradiated human allogeneic and autochthonous (autologous) tumor cells injected by the intralymphatic route. Immunoblotting studies on extracts of various established tumor cell cultures and fresh tumor biopsies were performed using sera from these patients. Evaluable tumor regressions were associated with detection of antibodies against human tumor cell antigens of 22,000 daltons (22 kd), 38,000 daltons (38 kd), 43,000 daltons (43 kd), and 70,000 daltons (70 kd). Similar antigens of approximately 22, 43, and 70 kd have also been detected in fresh extracts of certain human tumor tissues when tested with antisera from patients responding to immunotherapy. Production of antibodies to these antigens may play a role in tumor regression with active specific immunotherapy. These human regression-associated antigens may, therefore, represent novel agents for cancer immunotherapy.

Adult↗

Intralymphatic interleukin-2 in combination with zidovudine for the therapy of patients with AIDS.

In a pilot study the safety and therapeutic effects of an immunostimulatory intralymphatic treatment with natural human interleukin-2 (IL-2) in combination with zidovudine were evaluated in nine patients with AIDS. Therapy with IL-2 consisted of one subcutaneous injection of 0.1 microgram/kg IL-2, followed by four intralymphatic IL-2 infusions of 0.1 microgram/kg each within a period of up to 15 days. Enlargement of lymph nodes was seen in six and a transient increase of CD4 cells in five out of nine persons in association with the IL-2 therapy. An increase of HIV p24-antigenemia was observed only in the two patients in whom zidovudine dosage had to be reduced because of side effects. Moderate clinical side effects occurred in eight of the nine patients. Four patients developed zidovudine associated anemia. Six participants showed a favourable course of disease with survival of 25 to 54 months (median 30 months) despite a previous diagnosis of manifest AIDS before IL-2 therapy. This pilot study demonstrates that a combination therapy with intralymphatic IL-2 and zidovudine can induce positive immunomodulatory effects, even in the presence of manifest AIDS. Further studies should explore the tolerability and effects of a prolonged therapy with IL-2 in combination with a more potent antiviral drug combination therapy.

Acquired Immunodeficiency Syndrome↗

Serum levels of bleomycin released from the lymphatic system.

Endolymphatic injection Oil-Bleo (Nippon Kayaku, Japan) was performed on 24 patients for treatment of residual retroperitoneal malignant lymphoma detected after intravenous chemotherapy. The drug was administered once only in a dosage of 60 mg/30 mg per foot. Serum levels of the drug were monitored. The results were compared with data obtained after routine intravenous infusion of aqueous bleomycin to patients with advanced cervical cancer. The results revealed that bleomycin administered intralymphatically reaches maximum concentration after 15 minutes, and bleomycin after intravenous administration reaches maximum concentration at 36 hours.

Bleomycin↗

[Percutaneous injection of cisplatin lipiodol suspension (CLS) in rabbit lung, aiming at intratumoral injection therapy for lung cancer].

Cisplatin lipiodol suspension (CLS: cisplatin 20 mg/ml) was percutaneously injected (cisplatin dose, 2, 4 or 6 mg/kg) in normal lungs of 10 rabbits (1.9-2.3 kg) to assess the safety and feasibility of intratumoral injection of CLS for lung cancer. Histological study revealed acute and chronic infiltrates with bronchiolitis and immature fibrosis at the injected lung tissue even at four weeks after injection. Intrathoracic leaks of CLS produced mild and focal fibrinous pleuritis. Intrabronchial leaks of CLS produced peripheral bronchiolitis with regenerative epithelia. However, no noxious parenchymal damage in the lung and surrounding tissues was noted. Neither oil embolism in brain nor renal toxicity was demonstrated. Seven of eight rabbits showed an increase in body weight. Concentration levels of plasma platinum were lower when compared with intravenous injection of cisplatin in the rabbit: highest at 30 minutes and unmeasurable one week after injection. Lipiodol accumulation in mediastinal lymph nodes was demonstrated in two of nine rabbits by X-ray examination, suggesting intralymphatic drainage of CLS. Intratumoral injection of CLS is safe even with CLS leaks in surrounding normal lung tissues and may be a potent therapy for controlling mediastinal lymph nodes metastasized from lung cancer as well as the primary tumor.

Administration, Cutaneous↗

Radiocolloid lymphoscintigraphy in neoplastic disease.

The transport and intralymphatic deposition of interstitially injected radiocolloid of suitable physical properties are mediated through physiological processes and provide the means for obtaining scintigraphic images of drainage lymph nodes relevant to the injection site. In a large series of patients with breast carcinoma, high correlation has been shown between the internal mammary lymphoscintigram and clinicopathological stage of disease and prognosis. Since radiocolloid transport to, transit through, and deposition within the lymph node are effected through cellular elements concerned with immunological mechanisms, it is proposed that the radiocolloid lymphoscintigram be viewed not only as a technique for documenting morphologically established neoplasms in regional lymph nodes but also as a modality for the recognition of functional changes which may influence the development of such neoplasms.

Animals↗

Intralymphatic administration of liposome-encapsulated drugs to mice: possibility for suppression of the growth of tumor metastases in the lymph nodes.

The antimetastatic effects of two drugs, cis-diamminedichloroplatinum(II) (DDP) and hydrocortisone (liposome-incorporated or free), were studied. The experimental models were regional and distant metastases of hepatoma A and pulmonary adenocarcinoma, which were transplanted into the footpads of A/He mice. Liposomes were prepared from phosphatidylcholine by sonic dispersion. DDP and hydrocortisone were injected sc into the region of the plantar aponeurosis of the foot with the primary tumor. This administration route was considered to be equivalent to the intralymphatic route. Evidence indicated that only liposome-incorporated DDP and hydrocortisone decreased significantly the frequency and growth rate of tumor metastases in the regional lymph nodes. The effect observed was not due to the direct action of the drugs on the primary tumors. When nonencapsulated, these drugs were ineffective. Both liposome-encapsulated and free DDP did not affect distant metastases of pulmonary adenocarcinoma. The intralymphatic administration of liposome-encapsulated antitumor agents is suggested as a method for the prophylactic treatment of tumor metastases in the lymph nodes.

Animals↗

Enhanced anti-cancer effects of intralymphatic aclarubicin on distal lymph node metastases: quantitative evaluation using a new experimental model in mice.

The anti-cancer drug aclarubicin (2.0 mg/kg body weight) was injected into the left popliteal lymph node (the primary draining node of the foot-pad region) or into the tail vein, 8 days after a subcutaneous inoculation of 5 x 10(5) P388 leukemia cells/mouse in the left hind paw foot-pad of mouse (donor). During this time, metastases were established in the lower para-aortic nodes (the secondary draining nodes of this region). On day 10, the lower para-aortic nodes taken from each donor were transferred intraperitoneally to a normal mouse (recipient). From the recipients' survival time, the viable P388 leukemia cell number in the para-aortic nodes per donor mouse was estimated with a calibration line. The recipients' survival curve in the intralymphatic chemotherapy group was statistically significantly better than that in the intravenous chemotherapy group.

Aclarubicin↗

Intralymphatic BCG in the treatment of gynecologic malignancies: a phase I study.

Thirteen patients with a variety of advanced gynecologic malignancies were administered BCG via the dorsal lymphatics of the lower extremity in addition to standard accepted forms of therapy. Prolonged febrile courses, lymphangitis and suppurative adenitis were observed along the lymphatic pathway of the injected lower limbs. There was no correlation between reaction to a standard anergy panel and survival. There was also no correlation between reaction to a standard anergy panel and the inflammatory response to intralymphatic BCG (ILP-BCG). There was, however, a positive correlation between the inflammatory response to ILP-BCG and survival. Intralymphatic administration of immunostimulants may conceivably be of value as ancillary therapy for use in gynecologic malignancy. However, complications of this approach to immunotherapy are significant and the method should not be used until complications are decreased.

Adult↗

Lymphatic clearance of the human skin in patients with acute deep vein thrombosis using a novel fluorescent technique.

The purpose of this study was to investigate lymphatic clearance of the human skin in patients with acute deep thrombosis of the femoral vein. In 13 patients with deep vein thrombosis and no other cause for swelling of the limbs, lymphatic clearance of the skin at the foot was measured. Ten microliters of fluorescein isothiocyanatedextran 150,000 were injected intradermally and the fluorescent light intensity of the deposit measured 10 min and 24 hours after injection by window densitometry. In addition, intralymphatic pressure was measured by the servo-nulling system. The results were compared with a sex- and age-matched control group. Fluorescent light intensity decreased by 23.8 +/- 12.3 arbitrary units or by a factor of 1.8 +/- 0.5 in patients with DVT after 24 hours, which was significantly less than in healthy controls (33.7 +/- 8.9 arbitrary units or by factor 5.0 +/- 4.1, p < 0.013). Intralymphatic pressure was not different between the two groups. These results indicate that lymphatic clearance is significantly reduced in the acute phase of deep venous thrombosis.

Adult↗

Induction of canine in vitro reactivity to alloantigen following intralymphatic immunization.

An experimental model has been developed using the dog to study the induction of systemic cell-mediated immunity following intralymphatic immunization (ILI) with allogenic cells. As detected in one-way mixed lymphocyte cultures, blastogenically-reactive immune peripheral blood lymphocytes were observed after the third ILI with 10(7) cells. The in vitro reactivity was augmented by a fourth ILI to a node not previously injected indicating that a response in one node was followed by the trafficing of memory cells to other nodes. No immune PBL were detected after four ILI with lower doses of 10(3) cells. However, these dogs subsequently responded to a single injection of 10(7) cells with high levels of immune lymphocytes which were detectable for up to 24 days. Apparently, ILI with 10(3) or 10(5) cells, while insufficient to produce detectable levels of alloreactive lymphocytes were sufficient for lymphocyte priming. Results obtained with this model will aid in ongoing human trials of intralymphatic immunotherapy of malignant disease.

Animals↗