Enhanced antigen presenting and T cell functions during late-phase allergic responses in the lung.

BACKGROUND: Allergic inflammation is a common feature of asthma and may contribute to both development and perpetuation of disease. The interaction of antigen-presenting cells (APC) with sensitized helper T lymphocytes (TC) producing Th2 cytokines may determine the inflammatory response. Recruitment of APC and TC to the lung during allergic responses has been demonstrated, but functional studies in humans have been limited. OBJECTIVE: This study examined the function of APC and TC accumulating at sites of inflammation after segmental allergen challenge (SAC). METHODS: Fifteen allergic patients underwent SAC, and cells from bronchoalveolar lavage (BAL) were collected after 24 hours. APC and TC from the blood and BAL were purified based on expression of the monocyte marker, CD14; the plasmacytoid dendritic cell (pDC) marker, BDCA4, identifying neuropilin-1 (NRP1); and the helper T cell marker, CD4. Functional activity was assessed using allergen-induced T cell proliferation. Flow cytometry identified cells expressing CD14 and NRP1. RESULTS: SAC resulted in a 12-fold increase in mononuclear cells having the morphologic appearance of blood monocytes. Most of these cells co-expressed CD14 and NRP1. After saline challenge, BAL mononuclear cells demonstrated little APC function. Following SAC, BAL mononuclear cells showed function equal to pDC from blood and greater than blood monocytes. Purified NRP1+ cells from BAL had even greater function than pDC cells from blood (P = .008). Using consistent sources of APC, enhanced proliferation of TC from lung compared to blood was also demonstrated (P = .002). CONCLUSIONS: The marked increase in APC function for allergen-specific TC proliferation during allergic inflammation is largely due to the recruitment of monocytes and dendritic cells. There is also an enhanced response in the lung TC population, consistent with recruitment of allergen-specific T cells. Interactions between recruited APC and TC may occur as an early event promoting allergic airway inflammation.

Efficacy analysis of ultrasound-guided local injection of botulinum toxin type A treatment with orthopedic joint brace in patients with cervical dystonia.

OBJECTIVE: To study the efficacy of ultrasound-guided local injection of botulinum toxin type A (BTX-A) treatment with orthopedic joint brace in patients with cervical dystonia (CD). PATIENTS AND METHODS: A total of 105 patients with cervical dystonia were selected and randomly divided into medication treatment group (A group), botulinum toxin treatment group under the guidance of ultrasound treatment (B group) and botulinum toxin under the guidance of ultrasound treatment combined with orthopedic joint brace treatment group (C group). Tsui scale and Spitzer quality of life index was applied to evaluate the spasm and quality of life. The scores of Tsui scale and Spitzer quality of life index were compared after ultrasound-guided local treatment for one month, three months and six months. RESULTS: The difference in Tsui and Spitzer scores before and after the treatment of oral medications were not statistically significant (p > 0.05). Whereas, the differences in Tsui and Spitzer scores before and after the treatment between local injection of BTX-A treatment group and orthopedic joint brace combined with BTX-A injection group were statistically significant (p < 0.05). Also, the difference in Tsui and Spitzer scores of orthopedic joint brace combined with BTX-A injection group at 3 months, and 6 months were statistically significant compared to local injection of BTX-A treatment group (p < 0.05). CONCLUSIONS: Ultrasound-guided local injection of BTX-A combined with orthopedic brace could significantly reduce muscle tension and improve quality of life.

Efficiency of ultrasound and water capsule-guided local injection of botulinum toxin type A treatment on patients with facial spasm.

OBJECTIVE: To study the efficacy of ultrasound and water capsule-guided local injection of botulinum toxin type A (BTX-A) treatment on patients with facial spasm. PATIENTS AND METHODS: One hundred and fifty-seven cases of facial spasm were randomly divided into oral drug treatment group (group A) (78 cases) and ultrasound and water capsule-guided local injection of botulinum toxin type A treatment group (group B) (79 cases). Cohen, Acbert spasm strength grade scores in each case with facial spasm were recorded. Therapeutic effect, duration, significant efficiency, and muscle spasm strength were compared before and three after treatment. RESULTS: The muscle spasm strength showed no significant change in group A after the treatment. However, the muscle spasm strength was decreased significantly in group B after treatment (p < 0.01). CONCLUSIONS: Ultrasound and water capsule-guided local injection of botulinum toxin type A treatment is a safe, effective, and simple treatment for patients with facial spasm.

Ultrasound-guided botulinum toxin injections and EMG biofeedback therapy the lower limb muscle spasm after cerebral infarction.

OBJECTIVE: To evaluate the clinical efficacy under ultrasound-guided injection of botulinum toxin-A (BTX-A) and EMG biofeedback treatment of the lower limb muscle spasm after cerebral infarction. PATIENTS AND METHODS: Thirty-six cases of lower limb muscle spasm after cerebral infarction hemiplegia were randomly divided into two groups, the treatment group and the control group respectively including 18 cases. Both groups of patients were injected with BTX-A at different sites on spastic muscles. Twenty-four hours later, the treatment group was administered EMG biofeedback. Then, the modified Ashworth scoring was employed to observe the curative effect of the two groups. RESULTS: After six weeks' injection, the treatment group scored better than the control group (p < 0.05). CONCLUSIONS: Ultrasound-guided injection of botulinum toxin type A at various sites with EMG biofeedback treatment of the lower limb muscle spasms after cerebral infarction is efficient and conducive to the rehabilitation of patients' motor functions.

Association of vitamin D and antimicrobial peptide production during late-phase allergic responses in the lung.

BACKGROUND: Vitamin D may play important roles in regulating immune responses and in defence against infectious diseases by effects on both innate and adaptive immune responses. Little is known regarding activation of vitamin D within airway tissues and its relationship to inflammation and antimicrobial responses. OBJECTIVE: The objective of this study was to investigate the activation of vitamin D within the airways and to define relationships between vitamin D metabolites and measures of inflammatory and antimicrobial responses assessed by bronchoalveolar lavage (BAL) during late-phase responses following allergen challenge of allergic subjects. METHODS: Segmental allergen challenge was performed with saline and allergen in 16 adult allergic subjects. BAL was performed in both saline and allergen-challenged sites 20-24 h. after challenge. Following extraction from BAL fluids, levels of 25-hydroxy-vitamin D (25(OH)D) and 1,25-dihydroxy-vitamin D (1,25(OH)(2)D) were assayed by specific radioimmunoassays. The cleavage product of cathelicidin, LL-37, was assayed by ELISA. Cellular constituents and albumin were measured. RESULTS: Levels of vitamin D metabolites were increased in concentrated BAL fluids after allergen compared to saline challenge. Levels of 1,25(OH)(2)D increased from largely undetectable to 2.5 pm (median; range: 1-29.5; P = 0.005) while 25(OH)D increased from 3.2 (0.8-6.2) to 6.2 (1.5-184.9) nm (P = 0.0006). Levels of LL-37 increased from 2.1 (1.4-4.1) to 14.5 (2.2-106.7) ng/mL BAL (P = 0.0005). Levels of LL-37, 1,25(OH)(2)D, and 25(OH)D following allergen challenge were correlated with each other (P < 0.0001), cellular changes, and levels of albumin (P < 0.001). CONCLUSIONS AND CLINICAL RELEVANCE: Levels of vitamin D metabolites, particularly 1,25(OH)(2)D, were low within the airways and increased after allergen challenge. The increases correlated with the magnitude of inflammation and increases in cathelicidin. Normalization to albumin suggested plasma exudation as a mechanism for the increases. The findings support a role for vitamin D in allergic and innate immune responses in the lung.

Effects of adjuvant chemotherapy on bone marrow mesenchymal stem cells of colorectal cancer patients.

BACKGROUND: Chemotherapy damages the bone marrow and that is one of the most important problems in the treatment of malignancies, particularly colorectal cancer. The aim of the present study was to assess the effects of surgical adjuvant chemotherapy for CRC patients on human MSCs using an in vitro culture system. METHODS: The bone marrows of 43 CRC patients were harvested for separation and culture of MSC at pre- and post-chemotherapy. The number of colonies forming unit-fibroblast (CFU-F) was counted. The adhesive function of MSC was assayed and the growth of colony-forming unit-mixed hematopoietic cell (CFU-Mix) on the MSC layer was observed. The concentration of IL-6, SCF and FLT-T3 proteins in the MSC culture supernatant were also detected by ELISA assay. RESULTS: In the CRC patients with chemotherapy, we have demonstrated that the CFU-F exhibit significantly decreased. We also showed that the adhesive rate of bone marrow mesenchymal stem cell (BMSC) was significantly decreased. The growth of CFU-Mix on the MSC layer was inhibited. Most importantly, decreased CFU-F and the adhesive rate of BMSC were correlated significantly with decreased interleukins and stem-cell factor (IL-6, SCF and FLT-3L) expressions in the CRC patients after chemotherapy. CONCLUSION: Our results suggest that MSCs of CRC patients can be damaged by chemotherapy. Our data also indicates that the decreased expression of haematogenesis factors may play an important role in the pathogenesis. In the future, the MSC refused may have a potential clinical application in chemotherapeutically treated patients.

Lysophosphatidic acid is detectable in human bronchoalveolar lavage fluids at baseline and increased after segmental allergen challenge.

BACKGROUND: Lysophosphatidic acid (LPA) is a biologically active lysophospholipid and a component of normal plasma. LPA binds to receptors expressed on circulating and structural lung cells and affects cell growth and activation. Whether LPA is present in the lung has not been previously reported. OBJECTIVE: To develop an assay to measure LPA in bronchoalveolar lavage (BAL) fluids, and to study the association between LPA and allergic airway inflammation. METHODS: Seventeen allergic subjects underwent bronchoscopy and segmental allergen challenge, followed 18 h later by BAL. Supernatants were analysed for LPA content using liquid chromatography and mass spectroscopy. Expression of LPA receptors on primary bronchial epithelial cells was analysed by immunolabelling, and the effects of LPA on epithelial cell barrier function was investigated by measuring transepithelial resistance. RESULTS: LPA was detectable in BAL from control lung segments, and significantly increased 18 h after allergen challenge. Polyunsaturated species of LPA were especially increased following segmental allergen challenge. LPA levels did not strongly correlate with the number or percentages of eosinophils, neutrophils of lymphocytes, whereas MIP-3alpha (CCL20) levels correlated significantly with the allergen-driven influx of lymphocytes. The levels of LPA from control sites correlated inversely with BAL protein content, suggesting that LPA promoted epithelial barrier integrity at baseline. Experiments using primary human bronchial epithelial cells confirmed that LPA tightened the epithelial cell barrier. CONCLUSION: Lysophosphatidic acid is detectable in human BAL fluids at baseline and its expression increases during allergic inflammation. LPA does not appear to be a dominant chemoattractant for eosinophils or lymphocytes during allergic airway inflammation. In the absence of ongoing inflammation, LPA may promote epithelial barrier integrity.

Identification of a novel cytokine, ML-1, and its expression in subjects with asthma.

A novel gene, designated ML-1, was identified from a human genomic DNA clone and human T cell cDNA sequences. The second exon of ML-1 gene shares significant sequence identity with the gene encoding IL-17 (IL-17). ML-1 gene expression was up-regulated in activated PBMCs, CD4(+) T cells, allergen-specific Th0, Th1, and Th2 clones, activated basophils, and mast cells. Increased expression of the ML-1 gene, but not IL-17, was seen following allergen challenge in four asthmatic subjects, suggesting its role in allergic inflammatory responses. ML-1 from transiently transfected COS-7 cells was able to induce gene expression and protein production for IL-6 and IL-8 (at 10 ng/ml of ML-1: for IL-6, 599.6 +/- 19.1 pg/ml; for IL-8, 1724.2 +/- 132.9 pg/ml; and at 100 ng/ml of ML-1: for IL-6, 1005.3 +/- 55.6 pg/ml; for IL-8, 4371.4 +/- 280.5 pg/ml; p < 0.05 for both doses vs baseline) in primary bronchial epithelial (PBE) cells. Furthermore, increased expression of ICAM-1 was found in ML-1-stimulated PBE cells (mean fluorescence intensity (MFI) = 31.42 +/- 4.39 vs baseline, MFI = 12.26 +/- 1.77, p < 0.05), a functional feature distinct from IL-17 (MFI = 11.07 +/- 1.22). This effect was not inhibited by a saturating amount of IL-17. These findings demonstrate that ML-1 is a novel cytokine with a distinct function, and suggest a different receptor for ML-1 on PBE cells.

Glucocorticoid treatment increases inhibitory m(2) muscarinic receptor expression and function in the airways.

M(2) muscarinic receptors on parasympathetic nerve endings inhibit acetylcholine release in the airways. In this study, the effects of dexamethasone on M(2) receptors in vivo and in primary cultures of airway parasympathetic neurons were tested. Treating guinea pigs with dexamethasone (0.1 mg/kg, daily for 2 d) substantially increased inhibitory M(2) muscarinic receptor function, decreasing airway responsiveness to electrical stimulation of the vagi. At the same time, dexamethasone decreased the response to acetylcholine but not to methacholine, suggesting that cholinesterase activity was increased. When both cholinesterase and M(2) receptors were blocked (using physostigmine and gallamine, respectively) vagally induced bronchoconstriction was increased to control values. In primary cultures of airway parasympathetic neurons, dexamethasone significantly decreased the release of acetylcholine in response to electrical stimulation. Blocking inhibitory M(2) receptors using atropine (10(-5) M) increased acetylcholine release. After the M(2) receptors were blocked there was no difference in acetylcholine release between control and dexamethasone-treated cultures. M(2) receptor gene expression was increased by more than fivefold in dexamethasone-treated cultures. Immunostaining of dexamethasone-treated neurons demonstrated more intense staining. Thus, decreased vagally mediated reflex bronchoconstriction after glucocorticoid treatment may be the result on increased M(2) receptor expression and function as well as increased degradation of acetylcholine by cholinesterase.

Cyclophosphamide is associated with pulmonary function and survival benefit in patients with scleroderma and alveolitis.

BACKGROUND: Lung inflammation (alveolitis) may cause lung fibrosis in scleroderma. OBJECTIVE: To determine whether cyclophosphamide treatment is associated with retention of lung function and improved survival in scleroderma patients with alveolitis. DESIGN: Retrospective cohort study. SETTING: Johns Hopkins and University of Maryland Scleroderma Center. PATIENTS: 103 patients with scleroderma who had bronchoalveolar lavage or lung biopsy. INTERVENTION: Cyclophosphamide therapy. MEASUREMENTS: 1) Serial measurement of forced vital capacity (FVC) and carbon monoxide diffusing capacity and 2) survival. RESULTS: During a median follow-up of 13 months after bronchoalveolar lavage or biopsy, patients with alveolitis who did not receive cyclophosphamide therapy experienced a decrease in FVC (mean difference, -0.28 L [95% Cl, -0.41 to -0.16 L] and -7.1% of the predicted value [Cl, -10.9% to -4.0%]). Carbon monoxide diffusing capacity also decreased in these patients (mean difference, -3.3 x mmol min(-1) x kPa(-1) [Cl, -4.6 to -2.1 mmol x min(-1) x kPa(-1)] and -9.6% of the predicted value [Cl, -16.7% to -2.4%]). During a median follow-up of 16 months, patients with alveolitis who received cyclophosphamide were more likely to have a good outcome (stabilization or improvement) in FVC (relative risk, 2.5 [Cl, 1.5 to 4.1]) and diffusing capacity (relative risk, 1.5 [Cl, 1.0 to 2.2]). These patients also had improved survival; the median survival rate was 89% (25th, 75th percentiles, 84%, 94%) compared with 71% (25th, 75th percentiles, 55%, 86%) in untreated patients (P = 0.01, log-rank test). CONCLUSIONS: The presence of lung inflammation identifies patients with scleroderma who are more likely to have worsening lung function. Lung function outcomes and survival are improved in patients with alveolitis who receive cyclophosphamide.

Virus- and interferon-induced loss of inhibitory M2 muscarinic receptor function and gene expression in cultured airway parasympathetic neurons.

Viral infections increase vagally mediated reflex bronchoconstriction. Decreased function of inhibitory M2 muscarinic receptors on the parasympathetic nerve endings is likely to contribute to increased acetylcholine release. In this study, we used cultured airway parasympathetic neurons to determine the effects of parainfluenza virus and of interferon (IFN)-gamma on acetylcholine release, inhibitory M2 receptor function, and M2 receptor gene expression. In control cultures, electrically stimulated acetylcholine release increased when the inhibitory M2 receptors were blocked using atropine (10(-)5 M) and decreased when these receptors were stimulated using methacholine (10(-)5 M). Acetylcholine release was increased by viral infection and by treatment with IFN-gamma (300 U/ml). In these cells, atropine did not further potentiate, nor did methacholine inhibit, acetylcholine release, suggesting decreased inhibitory M2 receptor function and/or expression. Using a competitive reverse transcription-polymerase chain reaction method, we demonstrated that M2 receptor gene expression was decreased by more that an order of magnitude both by virus infection and by treatment with IFN. Thus, viral infections may increase vagally mediated bronchoconstriction both by directly inhibiting M2 receptor gene expression and by causing release of IFN-gamma which inhibits M2 receptor gene expression.

Cultures of airway parasympathetic nerves express functional M2 muscarinic receptors.

To study the control of acetylcholine release from airway parasympathetic neurons, primary cultures of these cells were established. Guinea pig tracheas were disaggregated with collagenase and plated onto matrigel-coated plates in medium that contained cytosine arabinoside to inhibit growth of dividing cells. Over 7 to 10 days neurites grow from the cell bodies, reaching a length of 2 mm. The vast majority of the cells in these cultures were neurons, as identified by morphology and staining with Neurotag and with antibody to neuron-specific antigen protein gene product 9.5. Cultured neurons contained acetylcholine, which was released by electrical field stimulation. Thus these were parasympathetic neurons. Staining with antibodies to M1, M2, and M4 muscarinic receptors revealed the presence of only M2 receptors. Likewise, reverse transcription-polymerase chain reaction using primers for M1, M2, and M4 muscarinic receptors revealed mRNA only for M2 receptors. Blocking these M2 receptors using atropine potentiated the stimulated release of acetylcholine, demonstrating that the M2 receptors inhibit acetylcholine release, as they have been shown to do in vivo. Thus airway parasympathetic neurons can be grown in culture, they retain the ability to synthesize and release acetylcholine, and they express functional inhibitory M2 muscarinic receptors.

Clonal diversity of IL-4 and IL-13 expression in human allergen-specific T lymphocytes.

The expression of IL-4 and IL-13 was analyzed in a panel of short ragweed allergen (Amb a 1)-specific T-cell clones from an allergic subject and a nonallergic individual. The T cells from the allergic subject showed a predominantly TH0 phenotype. The T cells from the nonallergic individual produced undetectable levels of IL-4 and high level of interferon-gamma, suggesting a TH1 cytokine profile. However, all T-cell clones showed significantly higher levels of IL-13 secretion than IL-4 secretion, and no quantitative correlation could be found between the levels of IL-4 and IL-13 in the clones tested. Furthermore, both cytokines showed similar kinetics of expression in antigen-induced steady-state messenger RNA. Finally, both cytokines were induced by stimulation of the cells with either ionomycin alone or with a combination of ionomycin and phorbol myristate acetate. These results demonstrate that there is a significant clonal diversity and quantitative difference in the levels of IL-4 and IL-13 expression in allergen-specific human T cells.

IL-13 expression at the sites of allergen challenge in patients with asthma.

Atopic asthma is characterized by inflammatory responses of the airway and is associated with up-regulation of Th2 cytokines, notably IL-4 and IL-5. A recently described human cytokine, IL-13, is a potent in vitro modulator of various cell types, including monocytes, B cells, and endothelial cells. Similar to IL-4, it is also involved in the induction of IgE synthesis. However, the in vivo expression and function of IL-13 and its relation to disease remain to be defined. Using a segmental allergen challenge model, we have examined the in vivo expression of IL-13 in the bronchoalveolar lavage (BAL) cells of atopic patients. We found a significant enhancement of both IL-13 transcripts and secreted proteins in the allergen-challenged BAL compared with the saline-challenged control sites of asthmatic and rhinitic patients. In contrast, the expression of IL-13 transcripts was not detected in the BAL of two normal subjects challenged with the same dose of ragweed allergen. The cellular source of IL-13 mRNA was identified in the mononuclear cell fraction of the allergen-challenged BAL. The allergen-induced quantitative differences in the level of transcripts were confirmed by competitive PCR assays. These results suggest that the significant increase in IL-13 in the allergen-challenged BAL is primarily from the mononuclear cells and is involved in the regulation of allergen-induced late phase inflammatory responses.

Detection of allergen- and mitogen-induced human cytokine transcripts using a competitive polymerase chain reaction.

Human cytokines, IL-4, IL-5, and IFN-gamma play an important role in the regulation of IgE synthesis and atopic diseases. In this communication, we describe the development of a quantitative assay of steady-state cytokine mRNAs (IL-4, IL-5, and IFN-gamma) from a variety of cell sources, including peripheral blood mononuclear cells (PBMCs) stimulated with either a mitogen (PHA) or ragweed pollen allergen extract, and cells from allergen-challenged inflammatory sites. Quantitative analysis of IL-5, IL-4 and IFN-gamma transcripts was achieved by a competitive reverse transcription-polymerase chain reaction (RT-PCR) technique using internal standard (IS) cRNAs in the presence of specific oligonucleotide primers. Each IS was generated from a plasmid vector containing the respective cytokine cDNA modified by insertion with an SV40-DNA fragment. Both test RNA and IS were reverse-transcribed and subjected to the 'competitive' PCR in the same tube. We first demonstrate the linearity and reproducibility of this technique; second, we apply this competitive PCR assay to analyze quantitatively the expression of IL-4, IL-5, and IFN-gamma transcripts in PBMCs before and after stimulation with PHA or crude ragweed allergen. Finally, we analyzed cells isolated from the lung lavage fluids of an atopic subject following allergen challenge, and showed a significant increase of IL-4 and IL-5 transcripts, but not IFN-gamma, in the allergen-challenged site when compared to the control. This technique of PCR quantitation provides an easy and efficient tool to study the expression of cytokine genes in allergic inflammatory diseases.

Analysis of cytokine transcripts in the bronchoalveolar lavage cells of patients with asthma.

A panel of steady-state cytokine mRNAs was analyzed in the bronchoalveolar lavage (BAL) cells from asthmatic subjects or patients challenged with ragweed allergen. This was achieved by combining both qualitative and quantitative assays using the reverse transcription-polymerase chain reaction (RT-PCR). Analysis of BAL cells from six mild allergic asthmatic and five nonasthmatic, nonallergic subjects showed no qualitative differences in the profile of cytokine mRNAs (including interleukin [IL]-1 beta, IL-2, IL-5, IL-6, IL-8, and granulocyte/macrophage colony-stimulating factor), except for tumor necrosis factor-alpha, which was detected in three out of six asthmatic BAL samples but in none of the controls. A key cytokine, IL-5, has been implicated in the pathogenesis of allergic inflammation through the recruitment of eosinophils. We found a significant enhancement of steady-state IL-5 transcripts in the BAL cells from allergen-challenged as compared with the saline-challenged control sites of four asthmatic patients; furthermore, the cellular source for IL-5 mRNA was identified in the mononuclear cell fraction, but not in the purified eosinophils, of the allergen-challenged BALs. These results suggest that the significant increase of IL-5 transcripts is primarily from the infiltrating mononuclear cells. Our study also demonstrates the power of qualitative and quantitative PCR analysis in determining the molecular basis of allergic inflammatory diseases.

Markers of multiple hematopoietic-cell lineages in multiple myeloma.

Multiple myeloma is considered a cancer of mature plasma cells. Recent studies, however, suggest the possible involvement of early B cells and the expression of myelomonocytic antigens by myeloma cells. Using flow cytometry, we searched for evidence of the expression of genes specific for different hematopoietic lineages by tumor cells in bone marrow aspirates from 27 patients with aneuploid multiple myeloma. In addition to features characteristic of myeloma cells, we found evidence of the frequent expression by myeloma tumor cells of the pre-B-cell antigen CALLA (common acute lymphocytic leukemia antigen) (in specimens from 58 percent of patients) and of megakaryocytic (88 percent), myelomonocytic (65 percent), and erythroid (39 percent) surface markers. The proportion of tumor cells expressing the different markers varied among patients, from 2 to 100 percent of recognizable tumor cells. We conclude that cells of multiple lineages are involved in myeloma--a finding that is consistent with the hypothesis that there is a common primary neoplastic lesion for all hematologic cancers.

P-glycoprotein expression in plasma-cell myeloma is associated with resistance to VAD.

Tumor cell-associated expression of multidrug resistance (MDR) was quantitated in 22 patients with DNA-aneuploid myeloma using 2-parameter flow cytometry with monoclonal antibody (MoAb) C-219 for the detection of cytoplasmic p-170 and propidium iodide for nuclear DNA content. The proportion of cells expressing p-170 and the intensity of p-170-related fluorescence were determined for each patient. Among the 14 patients treated with vincristine-adriamycin-dexamethasone (VAD), the proportion of p-170-positive cells distinguished sensitive from resistant disease (P less than .01). Among a subgroup of seven patients with MDR analysis available prior to VAD therapy, two subsequent nonresponders had high proportions of C-219-reactive cells. The presence de novo of high proportions of p-170-expressing cells in another still untreated patient and in a further individual with resistance to dexamethasone and interferon (not associated with MDR) warrants systematic analysis of p-170 expression prior to therapy to determine its clinical implications for response to MDR-associated drugs as combined in the VAD regimen. Concurrent MDR expression by aneuploid tumor cells and cells in the diploid subcompartment may represent involvement of diploid cells in the myeloma disease process.