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Improved radioimmunolocalization of human tumor xenografts following subcutaneous delivery of monoclonal antibodies.

The localization of a radiolabeled murine monoclonal antibody reactive with choriocarcinomas to human choriocarcinoma xenografts following intravenous and subcutaneous injection was evaluated by gamma scanning and tissue sampling. Tumor xenografts were established in the popliteal node region of athymic nude mice after repeated innoculations of the hind foot pads with BEWO choriocarcinoma cells. In dual label specific antibody studies, tumor/non tumor uptake ratios following subcutaneous (resulting in considerable intralymphatic uptake) injection of 131I-5F9.3 were significantly higher than those achieved post simultaneous intravenous injection of 125I-5F9.3. Double label experiments with 131I-5F9.3 and a nonspecific antibody, 125I-UPC-10, following subcutaneous injection, demonstrated that the high localization to popliteal region tumors was largely due to antibody specificity. Gamma scans following subcutaneous antibody administration of specific antibody to tumor bearing animals showed tumors soon after subcutaneous injection, at times earlier than those typically seen following intravenous delivery. Similar subcutaneous injections showed little normal nodal uptake in BALB/c control animals on gamma scans. No correlation was seen between tumor localization by specific antibody between the intravenous and intralymphatic routes, implying a difference in the mechanisms of tumor uptake of antibody by these two routes. The subcutaneous approach to antibody delivery offers advantages over intravenous delivery in tumors of human origin, including higher tumor/non tumor ratios and earlier imaging times. This was true even though these tumors were many times larger than normal lymph nodes. This subcutaneous delivery advantage should be exploitable in imaging nodal metastases of human tumors.

Animals

Intralymphatic antibiotic delivery for reducing acute prosthetic graft infection.

The lymphatic system has been implicated as a source of synthetic graft contamination when grafts are implanted in the presence of a distal septic focus. In previous studies, radical lymphatic excision and ligation were shown to reduce acute graft sepsis. However significant lymphedema precluded its clinical application. The present study was undertaken to evaluate methods for reducing acute graft sepsis while avoiding lymphatic obstructive complications. Twenty dogs were divided into one control and two experimental cohorts. Femoral interposition grafts were placed in each dog. A hind paw septic focus was introduced and therapy included a control (Group I--no therapy), intravenous antibiotics in Group II and intralymphatic antibiotics in Group III. Graft, blood and tissue cultures from each dog were taken at 48 hours. Lymphatic antibiotic therapy resulted in significantly improved graft culture results when compared to the control (p = 0.0003) and intravenously treated animals (p = 0.007). Blood cultures in the intralymphatically treated group were also significantly better (p = 0.003) than the control group.

Animals

[Intralymphatic B.C.G. infusions. Preliminary results (40 cases)].

A new route of administration for BCG aimed at stimulating immunity in cancer patients is presented. 40 patients received an intra-lymphatic infusion of BCG and in 20 lymphnode dissection was carried out for bacteriological and histological study. Details of technique and the results are given with regard to the effects of BCG on pelvic and pre-vertebral nodes after injection via a foot vessel.

BCG Vaccine

Regional intralymphatic infusion (ILI) of irradiated tumor cells with evidence of distant effects.

A case of canine lymphoma with radiographically-documented involvement of the para-aortic nodes is reported. Intralymphatic infusions (ILI) of cultured irradiated autochthonous tumor cells to remote lymph node bearing areas were associated with a dramatic initial shrinkage of the para-aortic lymphadenopathies. Three ILI timing schedules were used consecutively during a course of 10 treatments, allowing a comparison of responses in the same animal. The reported case suggests that a normal lymph node can be effectively "stimulated" by the same agent approximately every 3 weeks. A possible schedule for intralymphatic infusion is proposed for further investigation.

Animals

Immunosuppressive effect of intra-lymphatic irradiation.

The effect of intra-lymphatic administration of Lipiodol-l131 on the number of T and B lymphocytes, and on the in vivo response to common antigens and to DNCB, was investigated in uremic patients in the pre-transplant period. T lymphocytes have been detected by rosette formation with sheep erythrocytes (E) and B lymphocytes by rosette formation with erythrocytes sensitized with antibody and complement (EAC). After irradiation it was observed a lymphopenia due to a statistically significant decrease in the number of both T and B peripheral blood lymphocytes and a depletion of these cells in the irradiated lymph nodes. Their ability to give delayed hypersensitivity response to common antigens in vivo was lost after intralymphatic irradiation. Two patients were able to develop sensitization to DNCB and this reaction was supressed in one of them by irradiation. These effects of intralymphatic irradiation on the immune system favours further study of this method in clinical renal transplantation.

B-Lymphocytes

Intralymphatic interleukin-2 treatment of a hemophiliac AIDS patient with defective interleukin-2 production.

To improve immune functions in an interleukin-2 (IL-2) deficient hemophiliac AIDS patient suffering from severe Pneumocystis carinii pneumonia, treatment with IL-2 was started in addition to standard antimicrobial therapy. Highly purified IL-2 was administered subcutaneously and then repeatedly intralymphatically in a manner similar to pedal lymphography. No toxicity was observed. The patient temporarily improved clinically as well as with regard to immunological functions. Particularly the in vitro response to phytohemagglutinin (PHA) could partly be restored, and skin tests revealed improved response to recall antigens. These findings indicate that IL-2 can be administered safely and effectively by the intralymphatic route and may--in addition to antibiotics--be of value in AIDS patients with severe opportunistic infections.

Acquired Immunodeficiency Syndrome

IL-2/LAK cell treatment for advanced cancers with emphasis on a novel administration.

Interleukin-2 (IL-2) is a substance produced by activated blood cells called helper T-lymphocytes and has been shown to stimulate the body's immune system. IL-2 may cause certain tumors to regress when administered intravenously to laboratory animals and humans. Lymphokine activated killer (LAK) cells are white blood cells that have been stimulated with IL-2 in vitro. LAK cells are capable of killing tumor cells both in vitro and in vivo, especially when given along with IL-2. Although this form of treatment has been found to be effective in patients with certain cancers who no longer benefit from standard forms of therapy, the anti-cancer effects of IL-2/LAK cell treatment are limited by the serious, life-threatening side effects of high-dose intravenous administration, and by the high cost. A treatment program with low-dose, intralymphatically-administered LAK/IL-2 in patients with advanced cancer is a promising alternative which circumvents these major problems and concerns, while maintaining high response rates.

Blood Component Transfusion

[Lymphographically detectable long-term changes caused by endolymphatic radionuclide therapy (ELRT) in the infradiaphragmatic lymph system].

In sixteen patients who had been treated 42 months before on an average for malignant melanomas of the lower extremity (stage I) by a unilateral endolymphatic therapy with 32P/131J-Lipiodol UF, bipedal lymphography with oily contrast medium was performed. In consequence of the high-dosed intralymphatic radiotherapy the lymph nodes of the treated side are markedly reduced in size and number; fourteen out of the sixteen patients, however, showed lymph nodes of normal size and structure which had not been coloured during ELRT. On the basis of this phenomenon, a possible mechanism of formation of metastases in previously treated lymph nodes is discussed. The presentation and extent of radiogenic reactions in the lymph vessels correspond to the changes known for percutaneous irradiation.

Diaphragm

[Intralymphatic antibiotic therapy of inflammatory complications in colonic cancer].

In 18 patients with the inflammatory complications of colonic cancer before and after the operation, kanamycin, or one of the antibiotics of a cephalosporin group was administered into the lymphatic vessels of the upper third of a thigh at a daily dose of 20-25 mg/kg of body weight. After 3-4 days, the state of the patients considerably improved. All the patients underwent radical operations with primary restoration of intestinal continuity. There were no progression of purulent complications, postoperative lethality.

Adult

[Intralymphatic administration of open radionuclides in complex treatment of rheumatoid arthritis].

A new method was developed for the treatment of patients with rheumatic arthritis using therapeutic effect of radioactive colloid aurum in the lower limb lymphatic vessels. Altogether 50 patients with various degrees of activity and stages of disease were treated. A positive therapeutic effect was observed in 84%. The method of endolymphatic radiotherapy with radioactive colloid aurum made it possible to give up cytostatic and glucocorticoid therapy and to reduce a dosage of nonsteroid antiinflammatory drugs.

Adult

Intralymphatic autochthonous tumor cell vaccine in canine lymphoma.

Canine lymphoma is analogous to non-Hodgkin's lymphoma and is responsive to similar cytotoxic agents. The purpose of this study was to investigate a tumor cell vaccine delivered via peripheral lymphatics as maintenance therapy after induction of remission with chemotherapy. Thirty dogs with histologically confirmed lymphoma were treated with chemoimmunotherapy, and were induced into remission with combination chemotherapy, followed by intralymphatic (IL) autochthonous tumor cell vaccines as maintenance therapy. All dogs received two cycles of chemotherapy and at least three vaccine infusions. The median survival was 13 months with 11/30 (36%) still alive at 1-3 years follow-up. The median remission duration was 16 weeks while on IL vaccines alone. The survival time and remission duration is longer than previously reported in canine lymphoma.

Animals

[Radiation dose to the lung following endolymphatic radionuclide therapy of melanoma of the lower limbs].

The authors present 230 patients submitted to postoperative intralymphatic radiotherapy (ELRT) with radioactive Lipiodol UF because of a malignant melanoma of the inferior extremity. The incidence and volume of a possible invasion of the contrast medium into the lung and the resulting radiation exposure is indicated. Even if very low quantities of the contrast medium (3.5 ml and 7.0 ml, respectively) are used, only 22% of the cases show no pulmonary activity at all. Those patients who underwent bipedal ELRT showed more frequently higher radiation exposure. A correlation between lymph node weight and radiation exposure of the lung could not be demonstrated.

Combined Modality Therapy

Chemotherapy versus chemotherapy with intralymphatic tumor cell vaccine in canine lymphoma.

Fifty-eight dogs with lymphoma were treated with combination chemotherapy (vincristine, cyclophosphamide, L-asparaginase, and doxorubicin HCl [VCAA]) followed by intralymphatic autochthonous tumor cell vaccine (CI). Thirty dogs received chemotherapy alone (VCAA). There was no overall significant difference in survival times between the two groups, although there was a trend toward prolonged survival in the CI group. Asymptomatic dogs (Stage A) and dogs less than 7 years of age with Stage A disease treated with CI had significantly longer survival. Dogs treated with CI had a significantly longer first remission. Regardless of treatment group, male dogs had significantly longer remission times compared with female dogs.

Age Factors

[Intralymphatic drug therapy--a pathogenetically substantiated method of treatment in peritonitis].

The method of endolymphatic drug therapy (EDT) suggested by the authors has some advantages over the method of endoarterial therapy (which has proved to be effective in clinical practice) because it is aimed not only at control of the microbial flora, but also at correction of rheologic and microcirculatory disorders, improvement of the processes of tissue respiration and metabolism, stimulation of the intestine, prevention of the damaging effect of enzymes, toxins, and poisons on the organs and tissues of the body, and at correction of disorders of the immune status with promotion of the barrier function of the lymph nodes and the lymphatic system as a whole. EDT may be recommended for wide use in the clinic in the treatment of patients with peritonitis and other serious surgical infections unresponsive to the generally applied methods of management.

Adjuvants, Immunologic

A pilot study of intralymphatic interleukin-2. II. Clinical and biological effects.

Interleukin-2 (recombinant methionyl human interleukin-2 alanine 125; IL-2) was administered intralymphatically to 12 patients with advanced cancer in a phase I trial. Doses were administered once a week for 6 weeks in a dosage escalation schedule; patients were entered in four groups at successively higher starting dosages. Toxicity occurred in a profile similar to that seen with intravenous IL-2. The maximum tolerated dose with this route/schedule was 275,000 units/kg, a figure not higher than expected with intravenous administration. T1/2 alpha was prolonged to 54 min from the 13 min figure we obtained with IL-2 given intravenously. Granulocytosis and eosinophilia were seen, along with lymphocytosis following initial lymphopenia. Anti-IL-2 antibodies were seen in 42% of patients (compared to 16% with this agent given intravenously), suggesting increased immunogenicity of this route/schedule. No clinical response was achieved. Immunologic effects will be reported separately but are summarized.

Adolescent