Little is known about the plasma cell subpopulations in bone marrow. Bone marrow aspirates can only provide percentages of cell types and may not be representative. In this study, biopsies (Bouin's fixed, paraffin-embedded) from patients with a variety of haematological and non-haematological diseases (autoimmune 4, myeloid neoplasia 3, staging biopsies for Hodgkin's or non-Hodgkin's lymphomas – all normal – 5, anaemia 2, carcinoma – bone marrow unaffected – 2, miscellaneous 5) but not plasma cell dyscrasia were retrieved from files. Consecutive 3 µm sections were stained with polyclonal anti-κ, anti-λ, anti-γ, anti-α, anti-µ or anti-δ immunoglobulin (Ig) antisera and the reactions visualized using the avidin–biotin immunoperoxidase labelling method (Dako). All positive cells/section were counted in each section and the areas of bone marrow of the haematoxylin/eosin (H/E) stained sections were measured using a Vidas image analyser (Kontron Electronic) and Videoplan interactive software. The number of cells/mm2 positive with each light and heavy chain antibody was then calculated. The median bone marrow area of the biopsies was 6·1 mm2 (percentiles 4·4, 7·6) and the range was 1·3–13·4 mm2. Positive cells were spread reasonably uniformly throughout the sections apart from the expected concentration around blood vessels. The morphology of Ig µ-positive cells was indistinguishable from that of small plasma cells. The median (percentiles) and range of positive cells with each light and heavy chain antibody are shown in Table I. The κ/λ ratios of the absolute counts ranged between 1·2/1 and 3·4/1; median 1·9/1 (1·3/1, 1·8/1). This study has documented for the first time the range of absolute numbers of different plasma cell subsets in bone marrow biopsies from patients without plasma cell dyscrasia but with a spectrum of diseases commonly encountered in routine haematological practice. In a study of a similar group (n = 17) of patients, Thiele et al (1988) found a mean absolute plasma cell level of 87 (± 34)/mm2 in Giemsa-stained bone marrow sections. This compares well with the median total of 69 κ +ve plus λ +ve cells/mm2 demonstrated in our study. Our findings provide baseline values for comparison with individual plasma cell subsets of patients with plasma cell dyscrasia. In particular, knowledge of the upper limits of subsets encountered in diseases unrelated to plasma cell dyscrasia will enable diagnosis of low volume Igδ +ve or Igµ +ve disease to be made even when there is no overall increase of plasma cell or lymphoplasmacytoid cell numbers in Giemsa-stained sections or even perturbation of the overall κ:λ ratio. Bone marrow biopsies provide optimal material for measurement of absolute numbers of any cell type. Having a framework of bony trabeculae, shrinkage of tissue during fixation is not a problem. The presence of fibrosis and solid areas of tumour that are factors rendering the assessment of aspirated material uncertain either do not affect measurement of cell content or at least can be visualized as interfering factors. We suggest that there is new information to be gained from determination of absolute numbers, in particular plasma cell subsets in bone marrow biopsies.
Two monoclonal antibodies were applied to benign, dysplastic, and malignant human colorectal tissues using immunohistochemical techniques on formalin fixed paraffin embedded material. RAP-5 antibody is directed against a synthetic peptide, reflecting an amino acid sequence of the ras oncogene p21 protein product. Despite using several different techniques and antibody dilutions differential staining between the various epithelial populations was not obtained. RAP-5 also showed other tissue components such as plasma cells, histiocytes, fibroblasts, smooth muscle and vascular endothelium. CA19-9 antibody recognizes an epithelial surface carbohydrate antigen originally derived from a human colorectal carcinoma cell line: it did not stain normal colorectal mucosa or adenomatous polyps, but showed focal expression of variable strength in regenerative, dysplastic, and cancerous mucosa in ulcerative colitis, and in non-colitic colorectal carcinoma. Neither antibody was found to be a reliable marker of the evolution of malignant mucosal changes, although CA19-9 may be of limited use in confirming adenocarcinoma of gastrointestinal origin.
Ovarian serous carcinoma (OSC) is the most common ovarian epithelial malignancy. Recently, a dualistic pathway of ovarian serous carcinogenesis has been proposed based on morphologic observations and molecular genetic analysis. In this scheme, low-grade OSC arises in a stepwise fashion from a benign serous cystadenoma through a usual serous borderline tumor through a micropapillary variant of serous borderline tumor. In contrast, the more common high-grade OSC arises de novo from the ovarian surface epithelium or the epithelium of cortical inclusion cysts with an as yet unrecognized precursor lesion. Although the division of OSC into low- and high-grade variants is gaining greater acceptance, and although there is accumulating molecular genetic evidence for this, there is little published information regarding a comparison of protein expression between these two types of OSC. In this study, we have investigated the immunohistochemical expression of a wide range of proteins in cases of low-grade (n = 22) and high-grade (n = 47) OSC. Antibodies used were p53, MIB1, BCL2, WT1, HER-2/neu, C-KIT, osteopontin, and survivin. For all antibodies, except MIB1, cases were scored as 0 (negative or occasional positive cells), 1+ (<10% cells positive), 2+ (10%-25% cells positive), 3+ (26%-50% cells positive), 4+ (51%-75% cells positive) or 5+ (>75% cells positive). For MIB1, the percentage of positive nuclei was calculated. There was a statistically significant higher expression of p53, MIB1, BCL2, HER-2/neu, and C-KIT in high-grade compared with low-grade OSC (P < 0.05). Thirty of 47 (64%) cases of high-grade OSC exhibited 5+ staining with p53 compared with 4 of 22 (18%) low-grade neoplasms. Twelve of 47 (26%) high-grade OSCs exhibited 5+ staining with BCL2 compared with 1 of 22 (5%) low-grade OSCs. The mean MIB1 proliferative index in high-grade OSCs was 55.4% compared with 23.0% in low-grade OSCs. Virtually all cases of both low-grade and high-grade OSCs exhibited diffuse nuclear positivity with WT1 and diffuse cytoplasmic positivity with survivin. Osteopontin expression was variable with no significant difference in expression between low-grade and high-grade OSC. Although expression of both HER-2/neu and C-KIT was significantly higher in high-grade compared with low-grade OSC, only rare cases exhibited strong positivity with these antibodies, which could be of therapeutic value in individual cases, although this would require additional molecular investigations. The significant differences in protein expression between low-grade and high-grade OSC provides further support for a different underlying pathogenesis. In particular, the differences in p53 immunoreactivity are in keeping with the observation that p53 gene mutation is more common in high-grade than low-grade OSC.
While allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative option for many hematologic malignancies, complications such as graft-versus-host disease (GVHD) result in significant morbidity and mortality. Conventional approaches to manage GVHD, such as prophylaxis with immunosuppressive agents or T-cell depletion strategies, are limited by increases in graft failure, viral-associated lymphoproliferative disorders, and disease relapse. Here we present a novel strategy to reduce the rates of GVHD by programming mobilized peripheral blood ex vivo with a cocktail of small molecules prior to allo-HCT. An established xenogeneic mouse model was used to examine the potential of this cell programming strategy to reduce rates of GVHD. Sub-lethally irradiated NOD-scid IL2rγnull (NSG) mice were transplanted with human peripheral blood mononuclear cells (PBMC) pulse treated ex vivo with either vehicle or a cocktail of two small molecules (FT1050+FT4145). Recipients of pharmacologically programmed PBMCs had significantly lower GVHD scores, decreased levels of circulating IFN-ɣ and enhanced survival relative to recipients of vehicle PBMCs (p<0.0001, Mantel-Cox log rank). In addition to xenograft-GVHD studies, we explored the impact of this cell programming strategy in a murine model of GVL. Lethally irradiated BALB/c (H-2Kd) recipient mice were transplanted with either control or FT1050+FT4145 programmed C57BL/6 (H-2Kb) CD8+ T cells and T cell-depleted bone marrow. Prior to allo-HCT, recipients were injected with 2x104 luciferase-expressing A20 lymphoma cells (A20-luc). Bioluminescence imaging was used to monitor the tumor burden over a 28 day period post-HCT. Donor cells programmed with this small molecule cocktail significantly improved survival (p<0.001) while retaining GVL effects against the A20 lymphoma cells. Combined, these studies demonstrate that pharmacologic programming of hematopoietic cells with FT1050+FT4145 prior to allo-HCT may offer an innovative therapeutic approach to reduce rates of GVHD without compromising GVL activity.
Direct measurement of monoclonal plasma cell mass in bone marrow biopsies may be a useful parameter to establish in plasma cell dyscrasia. In this study monoclonal plasma cells/mm in light chain immunoglobulin immunostained archival bone marrow sections from 22 patients in whom a diagnosis of multiple myeloma (MM) had been excluded but who had monoclonal proteins were counted by two observers at light microscopic level. There was good correlation between the counts of the two observers. The levels of monoclonal plasma cells/mm in biopsies were not related to the % counts in the aspirates taken at the same time as the biopsies. Three of seven patients with biopsy levels in excess of the polyclonal levels in patients without plasma cell dyscrasia developed progressive MM within the observation time. Monoclonal plasma cell levels/mm of bone marrow biopsies can be measured and they provide a useful parameter for the assessment of patients with low volume plasma cell dyscrasia.
Patients with substance use disorder (SUD) rely upon urine drug testing to support treatment adherence and to mitigate relapse. Before the onset of coronavirus 2019 (COVID-19), the logistical challenges of randomized observed collections for urine drug testing for the patient were significant. During COVID-19, these barriers were often insurmountable. Since SUD patients represent a population at a higher risk for complications from COVID-19, an alternative strategy to support COVID-19 testing was urgently needed. We designed and deployed a telehealth-based solution in which patients could use mobile devices to connect with trained collection professionals to perform observed urine collections, often referred to a UA (urinalysis). The solution was designed with patient-centered best practices for telehealth, stigma prevention, trauma-informed, empathy and compassion, and to remove barriers to access to care. This approach demonstrated high patient satisfaction scores thereby proving that it is possible to provide urine collection services in the patient's home via a telehealth technology, while still upholding SUD testing integrity best practices. This study lays the path for a more patient-centered way to support this population.
Patients with substance use disorders (SUD) are at increased risk of both coronavirus disease-19 complications as well as exacerbations of their current conditions due to social distancing and isolation. Innovations that provide increased access to support substance use disorder patients may mitigate long-term sequelae associated with continued or renewed drug use. To improve patient access during the coronavirus disease-19 pandemic, we deployed a mobile unit to enable access to urine drug testing where needed for patients suffering from substance use disorder. Over a 3-week pilot program, 54 patients received urine drug testing across 5 providers and 8 zip codes. The mobile unit was cost-effective, demonstrating a volume-dependent 19% lower cost compared to pre-coronavirus disease-19 patient service centers in a similar geographic region. The mobile unit was well-received by patients and providers with an average of 9 out of 10 satisfaction scores and allowed for access to urine drug testing for 67% patients who would not have received testing during this time frame. No statistically significant differences were found in substance use positivity rates in comparison to pre-coronavirus disease findings; however, some shifts in use included higher rates of fentanyl and opioid positivity and reductions in tetrahydrocannabinol and cocaine use in the mobile collections setting. Deployment of mobile collection services during the coronavirus disease-19 pandemic has shown to be an effective mechanism for supporting patients suffering from substance use disorder, allowing for access to care of this often stigmatized, vulnerable population.
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