PHA Skin Test Research Articles

Combination Chemotherapy for Metastatic Hepatic Carcinoma and the Relation of Immune Status to the Response

Metastatic carcinoma presents a significant challenge in cancer treatment due to its aggressive nature and propensity for resistance to traditional therapeutic approaches. This systematic research paper delves into the development and evaluation of a novel combination chemotherapy regimen involving the use of 5-fluorouracil, cyclophosphamide & mitomycin C, and PHA skin test in the treatment of metastatic carcinoma, lymphomas, and breast cancer. The study aims to investigate the efficacy and safety profile of this combined chemotherapeutic approach, utilizing scientific methodology and analysis to assess its potential as a promising treatment option for patients with advanced cancer.

Effects of egg storage on hatchability, chick quality, performance and immunocompetence parameters of broiler chickens

Pre-incubation egg storage is a necessity for the poultry industry. This study evaluated the effects of pre-incubation storage length of broiler eggs on hatchability, 1-day-old chick quality, subsequent performance, and immunocompetence. To this end, a total of 360 hatching eggs were stored for 4, 12, or 16 d prior to incubation. Hatchability and chick quality were assessed at hatch, and growth performance and immunocompetence parameters were assessed during a 35 d rearing period. Hatchability of set and fertile eggs, and embryonic mortality, were not affected by egg storage. On the contrary, 1-day-old chick BW and length were linearly negatively correlated with egg storage length (P-linear < 0.05). Nevertheless, BW corrected for egg weight prior to setting was unaffected, and corrected chick length was positively affected by storage length. One-day-old chick Tona score, navel quality, and post-hatch growth performance (BW at 7 and 35 d, cumulative feed intake, and feed conversion ratio at 35 d) were unaffected by egg storage (P, P-linear > 0.05). Lymphoid organ weights at 2 and 35 d, the titre of maternal anti-NDV antibodies, most of the thymocyte subpopulations defined by CD3, CD4, and CD8 cell surface expression in the thymus of 2-d-old chicks, cellular responses to the PHA skin test, humoral responses to primary SRBC, and NDV immunizations were also not influenced by length of storage (P, P-linear > 0.05). On the contrary, the length of egg storage was found to negatively influence the abundance of CD3+ CD4− CD8− thymocytes that represent the majority of γδ-T cells in the thymus of 2-day-old chicks, as well as the humoral response to booster NDV immunization of the birds. In brief, pre-incubation storage of broiler hatching eggs for up to 16 d did not affect most developmental and growth parameters investigated, except for BW and length at hatch. Egg storage was found to suppress some aspects of the immunocompetence of the birds, particularly aspects of acquired immunity.

Genetic and environmental variation in condition, cutaneous immunity, and haematocrit in house wrens.

BackgroundLife-history studies of wild bird populations often focus on the relationship between an individual’s condition and its capacity to mount an immune response, as measured by a commonly-employed assay of cutaneous immunity, the PHA skin test. In addition, haematocrit, the packed cell volume in relation to total blood volume, is often measured as an indicator of physiological performance. A multi-year study of a wild population of house wrens has recently revealed that those exhibiting the highest condition and strongest PHA responses as nestlings are most likely to be recruited to the breeding population and to breed through two years of age; in contrast, intermediate haematocrit values result in the highest recruitment to the population. Selection theory would predict, therefore, that most of the underlying genetic variation in these traits should be exhausted resulting in low heritability, although such traits may also exhibit low heritability because of increased residual variance. Here, we examine the genetic and environmental variation in condition, cutaneous immunity, and haematocrit using an animal model based on a pedigree of approximately 2,800 house wrens.ResultsEnvironmental effects played a paramount role in shaping the expression of the fitness-related traits measured in this wild population, but two of them, condition and haematocrit, retained significant heritable variation. Condition was also positively correlated with both the PHA response and haematocrit, but in the absence of any significant genetic correlations, it appears that this covariance arises through parallel effects of the environment acting on this suite of traits.ConclusionsThe maintenance of genetic variation in different measures of condition appears to be a pervasive feature of wild bird populations, in contradiction of conventional selection theory. A major challenge in future studies will be to explain how such variation persists in the face of the directional selection acting on condition in house wrens and other species.

The PHA test as an indicator of phagocytic activity in a passerine bird.

Several techniques in ecological immunology have been used to assess bird immunocompetence thus providing useful information to understand the contribution of the immunological system in life-history decisions. The phytohaemagglutinin (PHA)-skin test has been the most widely employed technique being interpreted as the sole result of T lymphocytes proliferation and hence used to evaluate acquired immunological capacity. However, the presence of high numbers of phagocytic cells in the swelling point has cast some doubt about such an assumption. To address this issue, we collected blood from 14 days-old nestlings of spotless starling (Sturnus unicolor), administered subcutaneous PHA immediately after and then measured the swelling response 24 hours later. Differential counts of white blood cells suggested that an intense development of acquired immunological defences was taking place. The phagocytic activity of both heterophiles and monocytes was also very intense as it was the swelling response. Moreover, our results show, for the first time in birds, a positive relationship between the phagocytic activity of both kinds of cells and the swelling response. This broadens the significance of the PHA test from reflecting T lymphocytes proliferation -as previously proposed but still undetermined in vivo- to evaluate phagocytosis as well. In other words, our data suggest that the PHA swelling response may not be considered as the only consequence of processes of specific and induced immunity –T lymphocytes proliferation- but also of constitutive and nonspecific immunity –heterophiles and monocytes phagocytosis. We propose the extensive use of PHA-skin test as an optimal technique to assess immunocompetence.

Expression of Concern: The PHA-Skin Test Reflects Acquired T-Cell Mediated Immunocompetence in Birds

Expression of Concern: The PHA-Skin Test Reflects Acquired T-Cell Mediated Immunocompetence in Birds

Developmental toxicity in white leghorn chickens following in ovo exposure to perfluorooctane sulfonate (PFOS)

Studies show that perfluorinated compounds cause various toxicological effects; nevertheless, effects on immune function and developmental endpoints have not been addressed at length. This study examined the effects of perfluorooctane sulfonate (PFOS) in white leghorn hatchlings on various developmental, immunological, and clinical health parameters. In addition, serum PFOS concentrations were determined by LC/MS/MS. Embryonic day (ED) 0 eggs were injected with either safflower oil/10% DMSO (control, 0mg/kg egg wt) or PFOS in safflower oil/10% DMSO at 1, 2.5, or 5mg/kg egg wt, and the chicks were grown to post-hatch day (PHD) 14. Treatment with PFOS did not affect hatch rate. Following in ovo exposure chicks exhibited increases in spleen mass at all treatment levels, in liver mass at 2.5 and 5mg/kg egg wt, and in body length (crown-rump length) at the 5mg/kg treatment. Right wings were shorter in all treatments compared to control. Increases in the frequency of brain asymmetry were evident in all treatment groups. SRBC-specific immunoglobulin (IgM and IgY combined) titers were decreased significantly at all treatment levels, while plasma lysozyme activity was increased at all treatment levels. The PHA skin test response decreased in relation to increasing PFOS dose. Serum concentrations where significant immunological, morphological, and neurological effects were observed at the lowest dose (1mg/kg egg wt) averaged 154ng PFOS/g serum. These concentrations fall within environmental ranges reported in blood samples from wild caught avian species; thereby, verifying that the environmental egg concentrations used for the injections do indeed relate to serum levels in hatchlings that are also environmentally relevant. These data indicate that immune alterations and brain asymmetry can occur in birds following in ovo exposure to environmentally relevant concentrations of PFOS and demonstrates the need for further research on the developmental effects of perfluorinated compounds in various species.

The PHA Test Reflects Acquired T-Cell Mediated Immunocompetence in Birds

Backgroundcological immunology requires techniques to reliably measure immunocompetence in wild vertebrates. The PHA-skin test, involving subcutaneous injection of a mitogen (phytohemagglutinin, PHA) and measurement of subsequent swelling as a surrogate of T-cell mediated immunocompetence, has been the test of choice due to its practicality and ease of use in the field. However, mechanisms involved in local immunological and inflammatory processes provoked by PHA are poorly known, and its use and interpretation as an acquired immune response is currently debated.MethodologyHere, we present experimental work using a variety of parrot species, to ascertain whether PHA exposure produces larger secondary than primary responses as expected if the test reflects acquired immunocompetence. Moreover, we simultaneously quantified T-lymphocyte subsets (CD4+, CD5+ and CD8+) and plasma proteins circulating in the bloodstream, potentially involved in the immunological and inflammatory processes, through flow cytometry and electrophoresis.Principal FindingsOur results showed stronger responses after a second PHA injection, independent of species, time elapsed and changes in body mass of birds between first and second injections, thus supporting the adaptive nature of this immune response. Furthermore, the concomitant changes in the plasma concentrations of T-lymphocyte subsets and globulins indicate a causal link between the activation of the T-cell mediated immune system and local tissue swelling.Conclusions/SignificanceThese findings justify the widespread use of the PHA-skin test as a reliable evaluator of acquired T-cell mediated immunocompetence in diverse biological disciplines. Further experimental research should be aimed at evaluating the relative role of innate immunocompetence in wild conditions, where the access to dietary proteins varies more than in captivity, and to ascertain how PHA responses relate to particular host-parasite interactions.

Assessment of white blood cell phagocytosis as an immunological indicator of methylmercury exposure in birds.

White blood cell (WBC) phagocytosis was investigated as a potential immunological indicator of methylmercury (MeHg) exposure in birds. The assay was first assessed using chicken WBCs dosed with MeHg in vitro either in whole blood or as isolated cells and later using blood of wild common loons exposed in vivo to a range of dietary MeHg and having a range of blood-Hg concentrations. Whole blood and isolated WBCs from captive chickens were exposed to a range of MeHg concentrations for 3 h. After MeHg exposure, cells were incubated with fluorescent latex microbeads (diameter = 1.75 microm), fixed, and analyzed for size, complexity, and fluorescent intensity by flow cytometry. MeHg significantly depressed phagocytic activity when added to isolated WBCs at concentrations > 0.01 microg/ml, but not when added to whole blood up to 50 microg/ml. Similarly, no significant relationship between the concentration of Hg in whole blood and phagocytic capacity of WBCs in free-living loons was observed. Our results suggest that the phagocytosis assay, although rapid and convenient for use in field studies with wildlife species, is not a responsive immunological indicator of MeHg exposure at environmentally realistic concentrations of blood-Hg in wild loons. Assays that measure other immunologic endpoints (e.g., bacterial killing assay, PHA skin test, and mitogen-induced lymphocyte proliferation) should be assessed with respect to their ability to detect MeHg immunotoxicity in wild birds.

Therapeutic immunostimulating effects of plant mitogens exemplified by the L4 isolectin of PHA.

A sizable body of evidence has accumulated showing that the characteristics of certain plant mitogens should endow them with valuable immunomodulating effects. Immune stimulation would be the fundamental function operating through the broad consequences of nonspecific T cell activation following binding with their CD3 or CD2 molecules. Based on in vitro evidences that PHA, operating through the LDCC pathway, might kill any tumor target if it remains present in adequate concentration, the administration of mitogens for cancer therapy would be rational, and the same mechanism should also justify these agents for treatment of certain infections. Being nonerythroagglutinating, although leukoagglutinating in higher concentrations, PHA-L4 serves as a suitable model for immunostimulating activities of the mitogens that can be applied directly or as in vitro activators of adoptive leukocytes. The PHA skin test can be utilized to gauge the serum levels of inhibitory glycoproteins that become elevated in a number of the disorders treatable by the mitogen, thus facilitating necessary dosage modifications. While PHA is the only mitogenic lectin that has been tested clinically, Con A and PWM are the two most widely studied among the alternatives, with others also available and many undoubtedly awaiting discovery. The development of practical methods for producing industrial quantities of nonagglutinating mitogens in pure form should be the goal, and accomplishing the latter might also answer certain hypersensitivity concerns.

Prophylactic effect of bestatin on the onset of invasive mole--clinical and fundamental studies.

This study was performed to determine whether bestatin (Ubenimex) has clinical prophylactic effects on the onset of invasive mole and a direct inhibitory effect on the growth of hydatidiform molar cells. A total of 49 patients with hydatidiform mole treated at Nagoya University Hospital from 1984 to 1990 were randomly divided into two groups, a bestatin administered-group and a bestatin non-administered group. Patients in the bestatin group were given 30 mg of bestatin orally and daily for three months just after their molar deliveries. There was no significant difference in age, gravidity, parity and gestational weeks between the two groups. There was also no significant difference in the duration of human chorionic gonadotropin (hCG) negative conversion in patients without invasive mole between the two groups. However, the incidence of invasive mole in the bestatin group (2/25, 8%) was significantly lower than that of the non-bestatin group (7/24, 29.2%). Nevertheless, there was no significant difference between the two groups in such immunological parameters as PHA skin test, PPD skin test, PHA stimulation index (PHA-SI), white blood cell (WBC) count lymphocytes % per WBC, OKT 3% per lymphocytes, OKT 4% per lymphocytes, OKT4/OKT8 and Leu 11% per lymphocytes. In vitro studies were performed with primary cultured hydatidiform moles. The result was that bestatin inhibited the secretion of hCG and 3H-thymidine uptake of hydatidiform molar cells. Thus, a possibility was suggested that bestatin directly inhibits the growth of hydatidiform molar cells and prevents the onset of invasive mole.

Supplemental effect of retinoids on immume function in vitamin a deficient rats

To investigate the effect of retinoids on immune function, retinyl acetate(A), retinoic acid(RA) and methyl 5,6-epoxyretinoate(Epo-A) were injected to vitamin A deficient (A DEF) rats and their effects were compared with that of A DEF rats. PHA skin test was carried out during the period of retinoid injection(1 μg/day). After the retinoid injection of 100–500 μg i.m., blood was drawn for measuring serum protein fractions, and the spleen was investigated. Reactivity to PHA was increased after the injection of A, RA and Epo-A. Serum IgG levels were elevated in Epo-A group. Spleen weight and its mononuclear cell count increased after the injection of A, RA, and Epo-A. Histologically, the periarterial sheath was attenuated, packed with small lymphocytes, and the germinal center was rudimentary in A DEF rats. In the A and RA injected group, on the other hand, lymphatic nodules with well developed germinal centers were present and large, clear cells were found diffusely around the periarterial region. These findings suggest that A and RA may act to maintain well developed B cells in A DEF rats. Furthermore, it is suggested that Epo-A can also act to enhance the immune function of A DEF rats.

IMMUNOLOGIC STATUS AND IMMUNOLOGIC CHANGES BY TREATMENT IN BREAST CANCER PATIENTS

Immunologic status was investigated in 273 patients with breast diseases (209 cases of primary breast cancer, 32 of recurrent breast cancer and 32 of benign breast disease). Before start of treatment, we examined 14 immunologic parameters about all patients to know the immunologic status. Patients were divided into the following 4 groups : A) operation only (28 cases), B) operation + adjuvant chemotherapy (FEMP) (28 cases), C) operation + FEMP + Levamisole (28 cases), D) operation + FEMP + Krestin (28 cases).The following results were obtained.1. Cell-mediated immunity in breast cancer patients was kept on the same level as benign breast disease in Stage I & II, and fell down slightly in Stage III, and was impaired definitely in Stage IV & recurrence.2. Lymphocyte blastogenesis to PHA & PHA-S were the most reliable parameters related with the breast cancer clinical staging (U.I.C.C.) and PHA skin test, peripheral lymphocyte count and T cell count ranked next place.3. From the result of average of five parameters (peripheral lymphocyte count, T cell & B cell count and PHA & PHA-S blastogenesis), immune response in early stage (I & II) breast cancer was rather higher than in benign breast disease.4. Immunoglobulin levels (IgG, IgA, IgM) did not related with staging.5. Suppression of cell-mediated immunity by radical mastectomy was very slight.6. Immunosuppression by adjuvant chemotherapy (FEMP combination) was severe and continued for almost one month after the therapy, but when combined with Levamisole or Krestin the immunosuppressive effect was prevented and/or recovered earlier.

Non-Hodgkin's lymphomaの免疫機能的分類に関する研究

Twenty-six patients with non-Hodgkin's lymphoma (NHL) born in Ehime Prefecture were classified immunofunctionally by testing E, EA and EAC rosettes, surface immunoglobulin, Ia antigen and terminal trasferase (TdT) of lymphoma cells. Geographical distribution of the immunofunctional cell type, clinical stage and PPD and PHA skin test values was examined. According to cell type, there were those types of patients: 13 cases of the T-cell type (50%), 10 cases of the B-cell type (38%) and 3 cases of the Null-cell type (12%). There was no true histiocyte type. In the geographic study, the percentage of the T-cell type in the Matsuyama district was less than that in Kyushu island, but more than that in the northeast part of Japan. The number of T-cell type patients was much larger in the southwest part of the prefecture. B-cell type patients in stage I or II and T-cell type patients in stage III or IV were predominant. Rare cases of the T-cell type with EA or EAC rosetting were observed. Cells from Null-cell type patients and a T-cell type patient with diffuse lymphoblastic histology were TdT-positive, but others were negative. PPD and PHA skin tests showed a much weaker reaction in the T-cell type than in the B- and Null-cell types.

頭けい部悪性しゅよう患者の非特異的免疫能

We studied the following nonspecific humoral and cellular immunologicai factors in 240 patients with head and neck malignant tumor. On humoral factors; serum globulin, serum immunoglobulin, complement, CEA and serum immunosuppressive factor, on cellular factors; CellS of blood lymphocyte, percentage of T, B and IgG•FcR+T lymphocyte, skin reaction to PPD and PHA, MIF and PHA-induced lymphocyte transformation. These patients have already had abnormal immunological conditions compared with control non-malignant tumor subjects before treatment. Of humoral factors, serum α1, α2, γ globulin, complement and immunosuppressive factor were significantly elevated. Of the cellular factors, the number of blood lymphocyte was depressed. However, no significant difference was found in other factors.To elucidate the influence of histological character of tumor, stage of disease, age of patients and method of treatments on the immunological conditions, the patients were examined by the following five tests; serum α2 globulin, serum immunosuppressive factor, cells of blood lymphocyte, PHA skin test and PHA-induced lymphocyte transformation. 1) Patients with maxillary cancer or malignant lymphoma showed remarkably significant elevation of the humoral factors and depression of the cellular factors. 2) In the patients with advanced tumor, all humoral faceors were elevated and the cellular factors were depressed in the majority of patients. 3) Depressive tendency to the cellular factors was remarkably observed with aging. 4) Although the humoral factors were elevated, the cellular factors remained unchanged after the operation. While the humoral factors were unchanged, the cellular factors were depressed after the irradiation.

Adjuvant immunotherapy of carcinoma colli with levamisole: Prevention of immunological depression following surgical therapy and radiotherapy

Of 57 patients with carcinoma colli who underwent radical surgery followed by radiotherapy, 15 were treated with levamisole at a dose level of 150 mg a day for 3 consecutive days, every other week, starting 3 days before surgery. Another 17 patients received levamisole in the same manner except that the administration started simultaneously with post-operative radiotherapy. The immune competence of patients was determined in terms of immune globulin level, DNCB test, PHA skin test and lymphocyte blastogenic response to PHA, and was compared between the levamisole-treated group and the control group. Levamisole had little influence on immune globulin levels and skin reactions to PHA. The DNCB test after levamisole therapy showed no difference in the rate of positive response to DNCB between the levamisole group and the control group. Levamisole, when started before surgery, prevented the postoperative depression of lymphocyte responsiveness to to PHA and could more or less enhance the lymphocyte blast formation that had been depressed by radiotherapy.

Clinical studies on the correlation between immunologic status and total gastrectomy combined with splenectomy

The influence of splenectomy on the immunologic status of stomach cancer patients was studied. Eight gastric cancer patients having undergone total gastrectomy combined with splenectomy were evaluated immunologically. Peripheral lymphocyte counts were depressed and continued to be so in splenectomized patients. PHA-induced lymphocyte blastogenesis decreased rapidly after total gastrectomy with splenectomy and thereafter increased gradually. PHA skin test seemed to take almost the same time course. These recoveries to the normal range were observed 4 weeks after operation. In 10 other patients who had undergone total gastrectomy with splenectomy for gastric cancer and had been recurrence-free for more than 6 months, peripheral lymphocyte and T lymphocyte counts significantly increased compared with 7 cases having undergone total gastrectomy only, though no difference was seen in both PHA skin test and PHA-induced lymphocyte blastogenesis between the 2 groups.

IMMUNOCOMPETENCE OF PATIENTS WITH MALIGNANT LYMPHOMA

The immunocompetence of patients with malignant lymphoma was examined and compared to that of patients with cancer in head and neck. The following is a conclusion obtained.1) There was no aggravation of general condition in either patient before the treatment.2) Patients with malignant lymphoma did not show the elevation of serum Ig-G level and serum-inhibitory factors detected in patients with cancer. The increase of a2-globulin in patients with malignant lymphoma was not so significant as in patients with cancer. 3) The cellular immunocompetence in patients with malignant lymphoma was impaired more markedly than in patients with cancer. Number of peripheral lymphocyte was decreased much the same, but PPD skin test, PHA skin test and PHA-induced lymphocyte transformation test were suppressed more markedly in patients with malignant lymphoma.4) The depressed cellular immunocompetence in these two disease might be caused by different mechanism. It is strongly suggested that the humoral factors may play a role in patients with cancer and cellular factors in patients with malignant lymphoma for the depressed immunocompetence.5) In the treatment of patients with malignant lymphoma, follow-up examinations of immunocompetence are required and immunotherapy is more necessary than in patients with cancer.