PubMed Health⌕ Search

Biomedical subjects

M Yeh

Publications and source records attributed to M Yeh.

At least 55 records · Page 3Linked to original sources
PubMed1995

Return rates and outcomes from ethnicity-specific mental health programs in Los Angeles.

OBJECTIVES: The present study compared the return rate, length of treatment, and treatment outcome of ethnic minority adults who received services from ethnicity-specific or mainstream programs. METHODS: The sample consisted of 1516 African Americans, 1888 Asian Americans, and 1306 Mexican Americans who used 1 of 36 predominantly White (mainstream) or 18 ethnicity-specific mental health centers in Los Angeles County over a 6-year period. Predictor variables included type of program (ethnicity specific vs mainstream), disorder, ethnic match (whether or not clients had a therapist of the same ethnicity), gender, age, and Medi-Cal eligibility. The criterion variables were return after one session, total number of sessions, and treatment outcome. RESULTS: The study indicated that ethnic clients who attended ethnicity-specific programs had a higher return rate and stayed in the treatment longer than those using mainstream services. The data analyses were less clear cut when treatment outcome was examined. CONCLUSIONS: The findings support the notion that ethnicity-specific programs seem to increase the continued use of mental health services among ethnic minority groups.

Adult↗
PubMed1994

Development and hormonal modulation of postnatal expression of intestinal alkaline phosphatase mRNA species and their encoded isoenzymes.

In the rat, intestinal alkaline phosphatase (IAP) activity in the duodenum, but not jejunum, increases on day 22-24 after birth and exhibits higher activity hydrolysing phenyl phosphate (PhP) than beta-glycerophosphate (beta GP) [Moog and Yeh (1973) Comp. Biochem. Physiol. 44B, 657-666]. The mechanism underlying these developmental changes remains unknown. To define possible mechanisms, we have measured IAP activity and mRNA levels, and analysed IAP mRNA species and isoenzymes on postnatal days 12, 18, 24 and 32. Duodenal IAP activity and mRNA content were identical on postnatal days 12 and 18, but were 7-fold and 3-fold higher on day 24, respectively than on day 18. The increased IAP activity exhibited a high PhP/beta GP ratio and was accompanied by initial appearance of the 3.0 kb mRNA and 90 kDa isoenzyme. On day 32, duodenal IAP activity did not increase over the levels on day 24, whereas mRNA levels doubled. The lack of enzyme increase might be related in part to increased apical release, as luminal IAP activity increased from 2% of total mucosal IAP on days 12 and 18 to 7% and 14% on days 24 and 32 respectively. In the jejunum, IAP activity decreased postnatally, but mRNA content was unaltered; only the 2.7 kb mRNA and 65 kDa IAP isoenzyme were present. Administration of cortisone or cortisone+thyroxine induced simultaneous appearance of the duodenal 3.0 kb mRNA and 90 kDa isoenzyme with an increased PhP/beta GP ratio. Thus postnatal increase in duodenal IAP activity is related to the expression of a 90 kDa PhP-preferring isoenzyme encoded by the 3.0 kb mRNA. The low-PhP/beta GP-ratio 65 kDa isoenzyme is expressed in the duodenum and in the jejunum and is encoded by the 2.7 kb mRNA.

Alkaline Phosphatase↗
PubMed1994

Mechanism of the impaired T-cell proliferation in adult rats exposed to alcohol in utero.

Although attempts have been made to assess the effect of ethanol on the immune responses in individuals with fetal alcohol syndrome, there is no consensus as to the effect of ethanol on the immune system. Evidence that fetal alcohol-exposed (FAE) humans and animals have diminished proliferative response of T-cells to mitogenic lectins is well established. However, little is known about the mechanism of a toxic effect of ethanol on T-cell growth. Thus, a rat model was used to delineate the mode of ethanol action on T-cell proliferation. We found that the diminished T-cell proliferation in young adult FAE rats was due to a decreased responsiveness to interleukin 2 (IL2), but not to an impaired production of IL2 and expression of IL2 receptors (IL2R). Furthermore, the decreased proliferative response did not result from the presence of an excessive suppressor T-cell activity. Measurements of [Ca+2]i and T-cell proliferation were concurrently performed in batches of cells from the same animals. It was demonstrated that an increase in [Ca+2]i induced by Concanavalin A (Con A) in T-cells from FAE rats was not impaired, although the T-cell proliferation induced by Con A was significantly diminished. The results of the IL2-binding study showed that the Kd values and the number of both high- and low-affinity IL2R binding sites on the T-cells of FAE rats were comparable to those of pair-, or chow-fed rats. Finally, the results of the kinetics and rate of the internalization of IL2 showed that (1) the amount of the internalized IL2 was significantly reduced in T-cells from FAE rats, and (2) the half-time (t1/2) for dissociation of IL2 from the receptors in the T-cells from FAE rats was also greater than that of the control rats. These results taken together indicate that ethanol suppresses T-cell proliferation by interfering with events following the IL2-IL2R interaction.

Animals↗
PubMed1994

Update on current models of HIV-related neuronal injury: platelet-activating factor, arachidonic acid and nitric oxide.

This review aims to summarize recent work related to the pathogenesis and possible treatment of neuronal injury in the acquired immunodeficiency syndrome (AIDS), especially with reference to potential neurotoxic substances released by HIV-infected or gp120-stimulated macrophages/microglia. Approximately a third of adults and half of children with AIDS eventually suffer from neurological manifestations, including dysfunction of cognition, movement, and sensation. Among the various pathologies reported in brains of patients with AIDS is neuronal injury and loss. A paradox arises, however, because neurons themselves are for all intents and purposes not infected by human immunodeficiency virus type 1 (HIV-1). This paper reviews recent evidence suggesting that at least part of the neuronal injury observed in the brains of AIDS patients is related to excessive influx of Ca2+ after the release of potentially noxious substances from HIV-infected or gp120-stimulated macrophages/microglia. There is growing support for the existence of HIV- or immune-related toxins that lead indirectly to the injury or demise of neurons via a potentially complex web of interactions between macrophages (or microglia), astrocytes, and neurons. HIV-infected monocytoid cells (macrophages, microglia or monocytes), especially after interacting with astrocytes, secrete substances that potentially contribute to neurotoxicity. Not all of these substances are yet known, but they may include eicosanoids, i.e. arachidonic acid and its metabolites, as well as platelet-activating factor. Other candidate toxins include nitric oxide (NO.), superoxide anion (O2.-), and the N-methyl-D-aspartate (NMDA) agonist, cysteine. Similarly, macrophages activated by HIV-1 envelope protein gp120 also appear to release arachidonic acid and its metabolites, and cysteine.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS Dementia Complex↗
PubMed1994

Transcriptional regulation of intestinal hydrolase biosynthesis during postnatal development in rats.

Lactase-phlorizin hydrolase (LPH) and sucrase-isomaltase (SI) are intestine-specific microvillus membrane hydrolases whose specific activities demonstrate reciprocal regulation during development but whose mechanisms of regulation have not been fully defined. To investigate transcriptional control of these two proteins, the rat LPH and SI genes were cloned, and antisense probes for preprocessed mRNAs (pre-mRNAs) were developed from intron sequence. LPH mRNA, as measured by quantitative ribonuclease (RNase) protection assays, was abundant before weaning and decreased two- to fourfold during weaning, whereas SI mRNA was first detected 14 days after birth and increased rapidly to abundant levels by age 28 days. LPH and SI pre-mRNA levels paralleled those of their respective mRNAs. LPH transcriptional rate declined during weaning, whereas that of SI increased during this time as determined by RNase protection assays of pre-mRNAs and nuclear run-on assays. In the adult rat, LPH mRNA was restricted to the jejunum and proximal ileum, whereas SI mRNA was detected throughout the small intestine, a pattern regulated by transcriptional rate as confirmed by nuclear run-on assays. Lactase and sucrase specific activities correlated well with their respective protein and mRNA concentrations in all experiments. We conclude that gene transcription plays a major role in the developmental and horizontal regulation of LPH and SI biosynthesis and that these two genes are regulated differently in rat small intestine.

Aging↗
PubMed1994

Stress exacerbates neuron loss and cytoskeletal pathology in the hippocampus.

Glucocorticoids (GCs), the adrenal steroids secreted during stress, endanger the hippocampus, compromising its ability to survive neurological insults. GCs probably do so by disrupting energetics in the hippocampus, thus impairing its ability to contain damaging fluxes of excitatory amino acids and calcium. Superficially, these observations suggest that stress itself should also exacerbate the toxicity of neurological insults. However, most studies have involved unphysiologic GC manipulations, limiting speculations about the endangering effects of stress. In this study, rats were infused with the excitotoxin kainic acid (KA) after either having been adrenalectomized and replaced with a range of physiologic concentrations of GCs, or having been stressed intermittently. We observed that within the CA3 region, increasing CORT concentrations exacerbated the KA-induced neuron loss, the extent of tau immunoreactivity, and of spectrin proteolysis. The transitions from low to high basal GC concentrations and from high basal to stress GC values were both associated with significant exacerbation of neuron loss and tau immunoreactivity; the extent of spectrin proteolysis was less sensitive to increments in GCs. As would be expected from these data, exposure to intermittent stress prior to KA infusion also exacerbated neuron loss, tau immunoreactivity, and spectrin proteolysis in CA3. Thus, physiological elevations of GCs, and stress itself, can exacerbate hippocampal neuron loss and the attendant degenerative markers following an excitotoxic insult. Of significance, seizure and hypoxia-ischemia provoke considerable GC stress responses, which may thus worsen the resultant damage. Furthermore, a number of neuropsychiatric disorders, as well as aging, are associated with elevated basal GC concentrations, which may endanger the hippocampus in the event of neurological insult.

Animals↗
PubMed1993

Impaired cytosolic free calcium response in splenic T-cells from mice fed with ethanol-containing diet.

Calcium-dependent signal transduction pathways of T-cell proliferation have been extensively studied in the past years. However, little is known about effects of ethanol on the calcium-dependent signal transduction pathway in T-cell proliferation. Thus, a murine model was used to determine effects of ethanol in vivo on T-cell proliferation and the intracellular free calcium concentration [Ca2+]i in response to Concanavalin A (Con A) and recombinant IL2 (rIL2) in T-cells. Splenic cells from young C57BL/6 mice, that had been fed on 3 different diets (ethanol-, maltose substitute- and standard liquid-diet) for 7-8 weeks were tested for their proliferative responses to Con A and rIL2. Concurrently, measurement was also made of [Ca2+]i in the nylon-wool-enriched resting T-cells induced by Con A and in Con-A-activated blast T-cells induced by rIL2. Our results showed that [Ca2+]i increases were seen in the splenic T-cells from three different groups of mice following Con A, but not rIL2 stimulation. However, this increase was much smaller in the splenic T-cells from ethanol-fed mice as compared to mice on maltose- or standard-diet. Furthermore, we also demonstrated that the impaired [Ca2+]i increase was seen in the T-cells of the same ethanol-fed mice having decreased the proliferative response to Con A. This reduced proliferation did not result from the presence of excessive suppressor T-cell activity. Finally, we also demonstrated that both the number of IL2 binding sites/cell and the Kd values of the low- and high-affinity IL2R on the T-cells from ethanol-fed mice were unaltered. Because evidence indicates that (1) a normal level of [Ca2+]i increase is a prerequisite for the production of IL2 by mitogen-stimulated T-cells, and (2) T-cells from ethanol-fed mice have normal capacities to produce IL2 that is the crucial growth factor controlling T-cells to progress through the cell cycle, these lines of evidence taken together with the results of this study suggest that the impairment in [Ca2+]i increases in T-cells from ethanol-fed mice may not be the primary factor contributing to the diminished T-cell proliferation in the same mice.

Animals↗
PubMed1993

Use of pup in a cup model to study gastrointestinal development: interaction of nutrition and pituitary hormones.

The novel technique of artificial rearing (AR) of rat pups circumvents the difficulty of controlling diet composition and caloric intake. For studies of effects of nutrition and hormone interactions on gastrointestinal development, an appropriate experimental approach is to use AR rats whose corticosterone production is inhibited or abolished. Hypophysectomized (Hx) rats were used to examine whether growth retardation after Hx results from reduced caloric intake. Hx, sham-Hx and intact rats were isocalorically fed a cow-milk formula from day 12 to 18. Mother-fed (MF) Hx and intact rats were used as baseline controls. MF-Hx showed retarded intestinal growth, decreased body weight gain and reduced skeletal growth. In contrast, AR-Hx rats showed intestinal hypertrophy, normal body weight gain and reduced skeletal growth. Intestinal lactase activity remained higher in MF-Hx or AR-Hx rats than in control groups. AR-Hx rats showed no precocious increase of intestinal maltase and sucrase activity as did AR controls. Trace levels of serum growth hormone was detectable in MF-Hx but not in AR-Hx rats. We conclude that caloric intake can promote intestinal and somatic growth in the absence of the pituitary gland and pituitary hormones are required for skeletal growth and intestinal enzymic differentiation.

Animals↗
PubMed1992

"Locked-in syndrome" after intrathecal cytosine arabinoside therapy for malignant immunoblastic lymphoma.

A 22-year-old man with malignant immunoblastic lymphoma had "locked-in" syndrome within 48 hours of receiving a single (100 mg) dose of intrathecal cytosine arabinoside (ara-C) in conjunction with intravenous ara-C, cisplatin, and doxorubicin. Eight hours after therapy, the patient had central hypoventilation and blurred vision that progressed to blindness within 3 hours. During the next 10 hours, he became completely quadriplegic but remained intermittently alert and was able to respond to commands by eye or head movements. Radiographic studies showed necrosis of the medulla and swelling of the entire spinal cord. The patient persisted in a locked-in state until his death 3 weeks later, after removal of life support systems. Autopsy confirmed extensive necrosis of the lower medulla, optic chiasm, cranial nerves I and IV, and spinal cord. This case was unusual for its severity. The temporal relationship to ara-C instillation favors a toxic idiosyncratic response to chemotherapy. The authors advocate caution when bolus intrathecal and intravenous ara-C are administered to a patient within a short time of each other.

Adult↗
PubMed1992

[The establishment of human anti-tetanus toxoid hybridomas with in vitro immunized human tonsil cells].

Human tonsil cells in vitro immunized with tetanus toxoid were fused with human-mouse heteromyeloma line RF to generate human-mouse hybridomas. Hybridoma 891112-50 was cloned and 2 subclones (891112-50-3 and -4) were obtained. The secreted antibodies from the subclones were antigen specific, since they did not cross react with three irrelevant antigens (OVA, TCS and F gamma G). The hybridomas were quite stable. After 13 passages in tissue culture flasks, they still retained their antibody secreting ability. From flow cytometry analysis the subclone 50-3 was more stable than the subclone 50-4. The human immunoglobulin contained in supernatant collected during regular passages was equivalent to 69.6 micrograms/ml.

Animals↗
PubMed1991

Cortisone and thyroxine modulate intestinal lactase and sucrase mRNA levels and activities in the suckling rat.

Glucocorticoids and thyroxine modulate postnatal intestinal sucrase and lactase activities. Whether changes in enzyme activity are accompanied by changes in enzyme mRNA levels were determined in day 6 rats given thyroxine, cortisone, or thyroxine plus cortisone and killed 3 days later. Cortisone induced precocious expression of jejunal sucrase activity which was enhanced when cortisone plus thyroxine was administered; sucrase mRNA changed in parallel. Jejunal lactase activity was unaffected by thyroxine and was increased after cortisone, but not after thyroxine plus cortisone. Jejunal lactase mRNA levels increased equally after cortisone or after cortisone plus thyroxine. Thus, cortisone induces coordinated increases in sucrase and lactase activities and in corresponding mRNA levels. Thyroxine only enhances cortisone induced sucrase expression and antagonizes cortisone by depressing lactase activity post-translationally.

Animals↗
PubMed1991

Posttranslational cleavage of rat intestinal lactase occurs at the luminal side of the brush border membrane.

The intestinal sucrase-isomaltase precursor is cleaved at the brush border membrane by luminal proteases. Whether the lactase precursor also is cleaved by luminal proteases is uncertain. Lactase synthesis and processing was studied in 0- and 15-day-old rats after IP administration of [35S]methionine, and changes in precociously cortisone-induced sucrase-isomaltase were used as an internal control. Mucosal lactase and sucrase-isomaltase were separately immunoprecipitated and analyzed by autoradiography after electrophoresis. In both 0- and 15-day-old rats, mucosal lactase appeared as a 200K lactase precursor band at 30 minutes and as 200K and 225K lactase precursor bands at 60 minutes and was cleaved to form a 130K lactase band 120-240 minutes after labeling; sucrase-isomaltase similarly appeared as 210K and 220K bands at 30-60 minutes and was cleaved to form 140K I and 120K S subunits by 240 minutes in day 15 rats. To determine the role of luminal proteases, intestinal segments were isolated in situ and the luminal contents were flushed 30 minutes after labeling. Unflushed segments were used as controls. Only lactase precursor and sucrase-isomaltase precursor were present 240 minutes after labeling in flushed intestinal segments, but lactase precursor and sucrase-isomaltase precursor were cleaved in unflushed segments. Addition of trypsin or elastase into the lumen of flushed segments resulted in partial cleavage of lactase precursor but not of sucrase-isomaltase precursor. Luminal contents collected from the small intestine of day 15 rats 120 and 240 minutes after labeling showed 35S-labeled 130K and 80K polypeptides in lactase immunoprecipitates. It is concluded that intestinal lactase is synthesized as lactase precursor and transported to brush border membrane and cleaved by luminal proteases, and the amino end polypeptide cleaved from lactase precursor is released into the lumen.

Animals↗
PubMed1991

Fever response in elderly nursing home residents: are the older truly colder?

OBJECTIVE: To test the hypothesis that many nursing home residents with an apparently blunted fever response (maximum temperature less than 101 degrees F) may actually have a significant change in temperature (delta T greater than or equal to 2.4 degrees F) which is not recognized because of a low baseline temperature. DESIGN: Retrospective chart review for cases of infection that met specific criteria and for chart-recorded baseline and infection temperatures. Chart-recorded baseline temperatures were prospectively compared with re-measurement of morning temperatures. SETTING: Nursing Home Care Unit of the VAMC West Los Angeles. PATIENTS: Random review of 40 residents' charts resulted in the detection of 69 infections among 26 residents over a 20-month period. Fifty randomly selected residents prospectively underwent comparison of chart-determined and actual re-measurement of baseline temperatures. RESULTS: In 50 randomly selected residents, the mean oral baseline temperature of 97.4 +/- 0.2 (degrees F +/- SEM) closely approximated the mean nurse-recorded measures in the charts (97.6 +/- 0.1). Chart review detected 69 infections among 26 residents, with 53 episodes having a temperature recorded during the infection. The mean maximum temperature (Tmax) during an infection was 101.3 +/- 0.3 (degrees F +/- SEM) but 47% (25/53) of the episodes had a "blunted" fever response (Tmax less than 101 degrees F). Of the 25 "blunted" fevers (Tmax less than 101 degrees F), about one-fourth demonstrated an adequate change in temperature from baseline (delta T greater than or equal to 2.4 degrees F) but failed to reach 101 degrees F because of a low baseline. Most infections (89%) had a Tmax greater than 99 degrees F. CONCLUSION: Establishing a nursing home patient's basal temperature and monitoring for changes in temperature (delta T greater than 2.4 degrees F) and/or lowering the threshold for recognition of fevers (to 99 degrees or 100 degrees F) in nursing home residents with a change in function should assist in early recognition of infections.

Aged↗
PubMed1991

Thyroxine and cortisone cooperate to modulate postnatal intestinal enzyme differentiation in the rat.

Interactions of cortisone and thyroxine (T4) in modulating jejunal sucrase and lactase expression were studied in rats during early postnatal life. Cortisone (50 micrograms/g body wt) precociously induced sucrase activity in days 5-16 rats and enhanced activity thereafter until day 22. T4 (1 microgram/g) plus cortisone evoked greater sucrase expression in day 9 or younger rats. T4 did not induce sucrase expression until day 13. Lactase activity was enhanced in rats younger than day 9 by cortisone, and this effect was abolished when T4 was added. In days 19 and 22 rats, cortisone depressed lactase; with T4, lactase activity was further decreased. T4 alone did not suppress lactase activity until day 19. Quantitation of jejunal enzyme content showed that sucrase catalytic activity was higher in day 22 than 19 or younger rats and lower in rats given T4 than cortisone. In contrast, lactase activity remained constant in all animal groups. In vivo [35S]methionine-labeling studies using day 9 rats showed that cortisone induced de novo synthesis of sucrase and increased 35S incorporation into lactase. Cortisone plus T4 increased 35S incorporation into sucrase further and significantly increased 35S incorporation into lactase. We conclude that 1) cortisone and T4 cooperatively stimulate sucrase expression and reduce lactase activity during early postnatal life and 2) reduction in lactase activity accompanied by increase in lactase synthesis suggests that cortisone and T4 regulate lactase activity at posttranslational level.

Aging↗
PubMed1991

Intestinal lactase expression and epithelial cell transit in hormone-treated suckling rats.

Cortisone- and/or thyroxine (T4)-induced changes in jejunal lactase activity and epithelial cell migration were studied to determine the relationship of these two events. In suckling rats given a single dose of cortisone on day 6, jejunal lactase activity increased by 37% and cell turnover rate by 95% 3 days later, whereas T4 alone induced no changes. After cortisone plus T4, jejunal lactase activity decreased by 23% while cell turnover rate increased by 176%. Among all animal groups, the patterns of lactase expression along the crypt-villus (C-V) axis were similar, being low at the C-V junction, increasing to a high plateau at the mid- or third quarter villus level, and decreasing slightly at the villus tip. The calculated epithelial cell age at half maximum lactase expression in cortisone-treated and cortisone plus T4-treated rats was 30 and 53% younger than in control rats. Maximum lactase activity in villus cells was approximately 45% higher in cortisone-treated than in control or cortisone plus T4-treated rats. Parallel measurements of sucrase and lactase activities along C-V axis showed elevated lactase activity at higher villus positions than lead sucrase activity, suggesting that cortisone action occurs in villus cells. Thus jejunal lactase expression may be modulated by 1) adjusting villus cell age required for maximum expression, 2) altering the level of lactase activity in villus cells, and 3) changing the turnover rate of lactase containing epithelial cells.

Aging↗
PubMed1991

[Cellular aspects of in vitro induction of antibody responses of human cells].

An in vitro system for induction of antibody responses of human cells has been established in our lab. B cell enriched fractions from excised human tonsils or trauma spleen were cultured for 7-14 days with tetanus toxoid or HBsAg vaccine with or without human T cell conditioned medium (C. M.) or a mixture of low concentrations of PWM and LPS (MTG). Positive antibody responses could be detected in cultures. Cells taken from different culture periods were subjected to FACS analysis in order to expound cellular changes during antibody induction periods so as to improve the in vitro antibody induction system. The results were described as follows: 1. Variations in total percentages of T cells during culturing periods seemed to be related its initial percentages. Cells with bigger initial percentages tended to decrease first and finally maintained at about 30%. While cells with smaller initial percentages tended to increase and finally also maintained at 30%. 2. CD4+ Th cells and CD8+ Ts cells from tonsils and spleen behaved somewhat differently. In tonsil cell cultures the percentages of CD4+ cells were often bigger than the percentages of CD8+ cells throughout the culture period. However, the inverted proportions of CD4+/CD8+ were shown in spleen cell cultures, especially in the culture with C. M. The possible relationships between the variations in CD4+/CD8+ proportions described as above and the intensities of antibody responses were discussed. Additionally, adding 1-Leucine-Methyl Ester showed no effects either on CD8+ or CD4+ cell percentages. 3. B cell (SIg+) percentages in both tonsil and spleen cultures were quite stable throughout the culture period, about 60% of total cells. CD19, a marker of B cell, was only present in part of the cultured SIg+ cells. The significance of the variations in CD19+, SIg+ cells was unclear. CD5+ B cells were known as cells secreting autoantibodies. Our results showed that these cells consistently maintained a relatively low percentage in the whole antibody induction period. 4. The reasonableness standard we used for "gating" in FACS analysis was discussed.

Antibody Formation↗
PubMed1991

[The in vitro induction of antibody responses of human lymphocytes and its application to hybridoma production].

Sources of immunized lymphocytes constitute one of the main obstacles in the production of human monoclonal antibodies. We tried to get them through in vitro immunization. Cells from excised tonsils or trauma spleens were used for the induction of antibody responses in vitro. Antibodies to different antigens including sheep red blood cells, ovalbumin, tetanus toxoid, and hepatitis B surface antigens were induced in 7-14 days' cultures. Taking tetanus toxoid as antigen, we analysed the various factors required for antibody induction with statistics analysis, which included cell separation method, T cell conditioned medium, antigen dosage, serum content, and concentration of mitogen PWM and LPS. The results showed: (1) The cell separation method influenced the antibody production significantly in comparison with other factors. It signified that immune cells' combination was the most influential factor. (2) Serum also constituted quite important influencing factor especially in the later period of culture. However, it did not make much differences if it attended 10% or so. The antigens and mitogens tended to be used at low concentration. (3) Due to the significant variation among individuals and among different antigens, it is suggested to set up the culture system with some flexibility so as to adapt to the variation in cells and antigens from different sources. The present culture system we use includes nylon wall column separation of cells, suitable range of antigens (three doses instead of one), and either 10% T cell conditioned medium or a mixture of 1 microgram PWM/ ml + 0.1 microgram LPS/ml. The human B lymphocytes stimulated in vitro with tetanus toxoid were used for the construction of human hybridomas.

Animals↗