RESUMO
OBJECTIVE: To examine supernatants (SNs) of human squamous cell carcinoma of the head and neck cell lines for soluble tumor-derived factors capable of inducing activation and proliferation of human immune cells. DESIGN: The SN of squamous cell carcinoma of the head and neck cell line PCI-50 was cultured in serum-free medium and tested for the ability to induce expression of activation antigens, proliferation, cytotoxicity against tumor cell targets and cytokine production by purified human natural killer (NK) and CD4+ T cells. RESULTS: Supernatant of PCI-50 promoted expression of the following activation markers on NK and T cells: CD25 (interleukin-2R-alpha), HLA-DR (major histocompatibility complex class II), CD54 (ICAM-1), CD71 (transferrin receptor), and CD69 (activation-inducing molecule). In addition, SN induced and significantly sustained (P < .01) proliferation of human unseparated peripheral blood lymphocytes and NK or T cells in culture. Purified human NK or T cells cultured in the presence of the SN and IL-2 (120 IU/mL) had significantly higher antitumor cytotoxicity than that mediated by NK or T cells cultured in AIM-V medium and IL-2. The SN induced cytokine (interferon gamma, tumor necrosis factor alpha, interleukin-6) production in purified NK or T cells. When the SN was fractionated by molecular weight-based filtration into fractions greater and less than 30 kd, the growth- and cytotoxicity-promoting activities were consistently detectable in the greater than 30-kd fraction. CONCLUSIONS: Culture SN of squamous cell carcinoma of the head and neck cell lines contain a soluble factor(s) capable of activating NK and CD4+ T cells and of promoting growth and antitumor cytotoxicity of these lymphocyte subsets in vitro.
Assuntos
Antígenos CD , Linfócitos T CD4-Positivos/imunologia , Carcinoma de Células Escamosas/imunologia , Células Matadoras Naturais/imunologia , Antígenos de Diferenciação de Linfócitos T/imunologia , Divisão Celular/imunologia , Citocinas/imunologia , Citotoxicidade Imunológica/imunologia , Antígenos HLA-DR/imunologia , Neoplasias de Cabeça e Pescoço/imunologia , Humanos , Molécula 1 de Adesão Intercelular/imunologia , Interferon gama/imunologia , Interleucina-1/imunologia , Interleucina-2/imunologia , Interleucina-4/imunologia , Interleucina-6/imunologia , Lectinas Tipo C , Ativação Linfocitária/imunologia , Receptores da Transferrina/imunologia , Células Tumorais Cultivadas , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Human NK cells can be separated into two functionally distinct subpopulations based on the ability to rapidly respond to IL-2 by adherence to solid surfaces. To determine functions of the NK cell subsets in solid tumor tissues, adherent (A) and nonadherent (NA) NK cells were evaluated for their ability to infiltrate multicellular tumor spheroids in vitro, to kill carcinoma (CA) cell targets in these spheroids, and to mediate antitumor activity in vivo. A-NK cells were less cytolytic than NA-NK cells against CA targets in single cell suspensions or in monolayers. However, A-NK cells showed a significantly better ability than NA-NK cells to infiltrate tumor tissues and kill tumor cells in spheroids of human squamous cell CA of the head and neck or breast CA. Perilesional delivery of human A-NK cells and IL-2 resulted in regression of established human squamous cell carcinoma of the head and neck tumors growing subcutaneously in immunosuppressed nude mice. Similarly, in a xenograft model of human gastric CA metastatic to liver of nude mice, a single intrasplenic injection of A-NK cells in combination with i.p. infusions of IL-2 significantly reduced the number of established hepatic metastases (p < 0.007) and prolonged survival of the mice (p < 0.003). In contrast, NA-NK cells were ineffective in either of the in vivo xenograft tumor models. These findings demonstrate that A-NK cells represent a biologically unique and important subset of NK cells that, in contrast to the rest of NK cells, function as effector cells in solid tumor tissues and, consequently, have a great antitumor therapeutic potential.
Assuntos
Adenocarcinoma/terapia , Neoplasias da Mama/terapia , Carcinoma de Células Escamosas/terapia , Neoplasias de Cabeça e Pescoço/terapia , Imunoterapia Adotiva , Interleucina-2/farmacologia , Células Matadoras Ativadas por Linfocina/imunologia , Neoplasias Hepáticas/secundário , Subpopulações de Linfócitos/imunologia , Adenocarcinoma/imunologia , Animais , Neoplasias da Mama/imunologia , Carcinoma de Células Escamosas/imunologia , Adesão Celular , Movimento Celular , Testes Imunológicos de Citotoxicidade , Feminino , Neoplasias de Cabeça e Pescoço/imunologia , Humanos , Interleucina-2/uso terapêutico , Células Matadoras Ativadas por Linfocina/efeitos dos fármacos , Células Matadoras Ativadas por Linfocina/transplante , Neoplasias Hepáticas/terapia , Subpopulações de Linfócitos/efeitos dos fármacos , Subpopulações de Linfócitos/transplante , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Organoides , Neoplasias Gástricas/patologia , Transplante Heterólogo , Células Tumorais CultivadasRESUMO
The supernatant of a cell line of squamous cell carcinoma of the head and neck (SCCHN), PCI-50, was previously shown to induce activation, promote proliferation and increase antitumor cytotoxicity of freshly purified human natural killer (NK) cells and CD4+ T lymphocytes [Arch Otolaryngol Head Neck Surg (1994) in press]. This supernatant was found also to promote the growth of a variety of hematopoietic cell lines, including Jurkat, THP-1, K562, NK-92 or Epstein-Barr-virus-transformed B cell lines. The Jurkat cell line was selected as a reporter cell in an 18-h proliferation assay established to measure the growth-promoting activity of PCI-50 supernatant. The presence of soluble tumor-derived factors able to induce proliferation of Jurkat cells was demonstrated in the supernatant produced by several other SCCHN cell lines but not in that produced by a gastric cancer cell line (HR) or renal cell carcinoma line (5117G8). The growth-promoting PCI-50 supernatant was shown to contain 28 +/- 0.5 pg/ml interleukin-6 (IL-6) in vitro but was negative for interferon gamma, IL-1, IL-2, IL-4, tumor necrosis factor alpha, granulocyte/macrophage-colony-stimulating factor and IL-12. The addition of any of these recombinant cytokines to Jurkat cell cultures did not significantly promote growth, while PCI-50 supernatant was consistently growth-stimulatory. This supernatant neither enhanced intracellular Ca2+ concentration in Jurkat cells nor induced up-regulation of activation antigens on the cell surface, although it supported growth of Jurkat cells in the absence of IL-2. The growth-promoting activity in the PCI-50 supernatant was acid-labile at pH 2 for 4 h, heat-resistant at 96 degrees C for 1 h and sensitive to treatments with trypsin and pepsin. Preincubation of the PCI-50 producer cells with tunicamycin or cyclohexamide reduced the level of growth-promoting activity in the supernatant. A partial purification of this activity was achieved using Amicon filtration, chromatography on concanavalin-A-Sepharose and then a hydroxyapatite column and high-pressure liquid chromatography gel filtration. The partially purified glycoprotein had a molecular mass of 50-70 kDa, as determined by gel filtration.
Assuntos
Carcinoma de Células Escamosas/patologia , Substâncias de Crescimento/farmacologia , Neoplasias de Cabeça e Pescoço/patologia , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Citocinas/farmacologia , Substâncias de Crescimento/isolamento & purificação , Humanos , Linfócitos/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Células Tumorais CultivadasRESUMO
OBJECTIVE: Human squamous cell carcinomas of the head and neck (SCCHN) have been shown to express interleukin 2 receptor (IL-2R), and binding of the ligand, IL-2, to the receptor results in tumor growth inhibition in vitro or in vivo in an SCCHN xenograft model in nude mice. To optimize growth inhibitory effects of IL-2, expression of the alpha or gamma chains of IL-2R in SCCHN was experimentally modified by transfection of tumor cells with the respective IL-2R genes or the lacZ gene as control. DESIGN: Using plasmid vectors containing the IL-2R alpha chain gene under the control of a cytomegalovirus promoter or the IL-2R gamma chain gene under the control of a Rous sarcoma virus promoter, the IL-2R genes were transferred by lipofection into SCCHN cell lines. Stable transfectants were selected, cloned by limiting dilution, and clones were compared with the parental cell lines for their sensitivity to the growth-inhibitory effect of IL-2. RESULTS: Transfer of the IL-2R alpha chain gene into SCCHN cells resulted in significant upregulation of expression of the IL-2R alpha chain on tumor cell surface but not in increased tumor growth inhibition by IL-2. In contrast, SCCHN IL-2R gamma transfectants, which expressed IL-2R gamma chain transcripts as confirmed in RNase protection assays, were significantly inhibited in growth and were sensitive to lower concentrations of IL-2 than the parental cell lines. CONCLUSIONS: Genetic modification of IL-2R expression on IL-2R-positive tumor cells in culture significantly alters their proliferative response to IL-2. These observations open a way for developing new strategies for therapy of SCCHN based on direct interactions of IL-2 with its receptor on tumor cells.
Assuntos
Carcinoma de Células Escamosas/genética , Regulação Neoplásica da Expressão Gênica/genética , Técnicas de Transferência de Genes , Neoplasias de Cabeça e Pescoço/genética , Receptores de Interleucina-2/genética , Divisão Celular/efeitos dos fármacos , Divisão Celular/genética , Depressão Química , Relação Dose-Resposta a Droga , Citometria de Fluxo , Humanos , Interleucina-2/farmacologia , Plasmídeos/genética , Transfecção/genética , Transfecção/métodos , Células Tumorais Cultivadas/citologia , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
The anatomy of the facial nerve relative to its intratemporal and extratemporal courses varies over time with developmental changes. Otologic and parotid surgery in infants and children demands detailed knowledge of the precise anatomy of the facial nerve with respect to the tympanic ring and external auditory canal. The authors analyzed this area using our three-dimensional (3-D) computer-aided reconstruction and measurement method studying the spatial relations of the facial nerve to the tympanic ring and stylomastoid foramen. Temporal bones from five normal individuals aged 36 gestational weeks, 3 months, 8 months, 4 years, and 17 years were retrieved from the temporal bone collection stored at the Elizabeth McCullough Knowles Otopathology Laboratory in Pittsburgh. Three-dimensional reconstruction of the facial nerve comparing the developmental anatomy across the various age groups provides the surgeon with the technical information necessary to address problems in this area.
Assuntos
Nervo Facial/anatomia & histologia , Processamento de Imagem Assistida por Computador , Processo Mastoide/inervação , Adolescente , Envelhecimento , Pré-Escolar , Nervo da Corda do Tímpano/anatomia & histologia , Desenho Assistido por Computador , Meato Acústico Externo/inervação , Orelha Média/inervação , Nervo Facial/embriologia , Feminino , Feto , Humanos , Lactente , Masculino , Osso Temporal/inervaçãoAssuntos
Proteínas de Ligação ao Cálcio/sangue , Osteogênese Imperfeita/sangue , Adolescente , Fosfatase Alcalina/sangue , Osso e Ossos/análise , Cálcio/sangue , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Minerais/análise , Osteocalcina , Osteogênese Imperfeita/metabolismo , Fósforo/sangueRESUMO
We evaluated the long-term use of synthetic salmon calcitonin in the management of osteogenesis imperfecta tarda and congenita. Forty-eight children, ranging in age from 6 months to 15 years, and two young adults, received synthetic salmon calcitonin 2 MRC units/kg three days a week and a daily oral calcium supplement of 230 to 345 mg. The annual fracture rate was decreased during calcitonin therapy as compared to the period preceding therapy. There was an increase in the ability of the patient to stand and move and in the subjective feeling of strength in the lower extremities during calcitonin therapy. There was also a significant improvement in radiographic bone density, as determined by the method of photodensitometry, in patients under 5 years of age. Long-term administration of synthetic salmon calcitonin may be beneficial to young children with osteogenesis imperfecta.