Use of a whole blood assay to monitor the immune response to mycobacterial antigens in leprosy patients: a predictor for type 1 reaction onset?

The stable incidence of new leprosy cases suggests that transmission of infection continues despite worldwide implementation of MDT. Thus, specific tools are needed to diagnose early stageMycobacterium lepraeinfection, the likely sources of transmission.M. lepraeantigens that induce T-cell responses inM. lepraeexposed and/or infected individuals thus are major targets for new diagnostic tools. Previously, we showed that ML1601c was immunogenic in patients and healthy household contacts (HHC). However, some endemic controls (EC) also recognized this protein. To improve the diagnostic potential, IFN-γresponses to ML1601c peptides were assessed using PBMC from Brazilian leprosy patients and EC. Five ML1601c peptides only induced IFN-γin patients and HHC. Moreover, 24-hour whole-blood assay (WBA), two ML1601c peptides could assess the level ofM. lepraeexposure in Ethiopian EC. Beside IFN-γ, also IP-10, IL-6, IL-1β, TNF-α, and MCP-1 were increased in EC from areas with high leprosy prevalence in response to these ML1601c peptides. Thus, ML1601c peptides may be useful for differentiatingM. lepraeexposed or infected individuals and can also be used to indicate the magnitude ofM. lepraetransmission even in the context of various HLA alleles as present in these different genetic backgrounds.

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Assessment of immune response to meningococcal disease: comparison of a whole-blood assay and the serum bactericidal assay

Microbial Pathogenesis ◽

10.1006/mpat.1999.0296 ◽

1999 ◽

Vol 27 (4) ◽

pp. 207-214 ◽

Cited By ~ 27

Author(s):

C.A Ison ◽

N Anwar ◽

M.J Cole ◽

R Galassini ◽

R.S Heyderman ◽

...

Keyword(s):

Immune Response ◽

Meningococcal Disease ◽

Whole Blood ◽

Whole Blood Assay ◽

Bactericidal Assay ◽

Blood Assay ◽

Serum Bactericidal Assay

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Use of a whole blood assay to evaluate in vitro T cell responses to new leprosy skin test antigens in leprosy patients and healthy subjects

Clinical & Experimental Immunology ◽

10.1046/j.1365-2249.1999.00892.x ◽

1999 ◽

Vol 116 (2) ◽

pp. 263-269 ◽

Cited By ~ 29

Author(s):

WEIR ◽

BRENNAN ◽

BUTLIN ◽

DOCKRELL

Keyword(s):

T Cell ◽

Skin Test ◽

Whole Blood ◽

Healthy Subjects ◽

T Cell Responses ◽

Whole Blood Assay ◽

Cell Responses ◽

Blood Assay ◽

Leprosy Patients

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Stimulated Immune Response by TruCulture® Whole Blood Assay in Patients With European Lyme Neuroborreliosis: A Prospective Cohort Study

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.666037 ◽

2021 ◽

Vol 11 ◽

Author(s):

Mathilde Ørbæk ◽

Rosa Maja Møhring Gynthersen ◽

Helene Mens ◽

Christian Stenør ◽

Lothar Wiese ◽

...

Keyword(s):

Immune Response ◽

Cohort Study ◽

Whole Blood ◽

Clinical Manifestations ◽

Lyme Neuroborreliosis ◽

Symptomatic Infection ◽

Whole Blood Assay ◽

Th17 Response ◽

Blood Assay ◽

Ifn Γ

IntroductionBorrelia burgdorferi sensu lato complex (B. burgdorferi) can cause a variety of clinical manifestations including Lyme neuroborreliosis. Following the tick-borne transmission, B. burgdorferi initially evade immune responses, later symptomatic infection is associated with occurrence of specific antibody responses. We hypothesized that B. burgdorferi induce immune hyporesponsiveness or immune suppression and aimed to investigate patients with Lyme neuroborreliosis ability to respond to immune stimulation.MethodsAn observational cohort study investigating the stimulated immune response by standardized whole blood assay (TruCulture®) in adult patients with Lyme neuroborreliosis included at time of diagnosis from 01.09.2018-31.07.2020. Reference intervals were based on a 5-95% range of cytokine concentrations from healthy individuals (n = 32). Patients with Lyme neuroborreliosis and references were compared using Mann-Whitney U test. Heatmaps of cytokine responses were generated using the webtool Clustvis.ResultsIn total, 22 patients with Lyme neuroborreliosis (19 definite, 3 probable) were included. In the unstimulated samples, the concentrations of cytokines in patients with Lyme neuroborreliosis were comparable with references, except interferon (IFN)-α, interleukin (IL)-17A, IL-1β and IL-8, which were all significantly below the references. Patients with Lyme neuroborreliosis had similar concentrations of most cytokines in all stimulations compared with references. IFN-α, IFN-γ, IL-12 and IL-17A were lower than references in multiple stimulations.ConclusionIn this exploratory cohort study, we found lower or similar concentrations of circulating cytokines in blood from patients with Lyme neuroborreliosis at time of diagnosis compared with references. The stimulated cytokine release in blood from patients with Lyme neuroborreliosis was in general slightly lower than in the references. Specific patterns of low IL-12 and IFN-γ indicated low Th1-response and low concentrations of IL-17A did not support a strong Th17 response. Our results suggest that patients with Lyme neuroborreliosis elicit a slightly suppressed or impaired immune response for the investigated stimulations, however, whether the response normalizes remains unanswered.

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Host immune responses induced by specific Mycobacterium leprae antigens in an overnight whole-blood assay correlate with the diagnosis of paucibacillary leprosy patients in China

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0007318 ◽

2019 ◽

Vol 13 (4) ◽

pp. e0007318

Author(s):

Xiaohua Chen ◽

Yuan-Gang You ◽

You-Hua Yuan ◽

Lian C. Yuan ◽

Yan Wen

Keyword(s):

Immune Responses ◽

Whole Blood ◽

Mycobacterium Leprae ◽

Whole Blood Assay ◽

Host Immune Responses ◽

Blood Assay ◽

Leprosy Patients

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Non-Thrombogenicity of Clean Glass Revealed by Native Whole Blood Assay in Bead Columns

Thrombosis and Haemostasis ◽

10.1055/s-0038-1665319 ◽

1983 ◽

Vol 50 (04) ◽

pp. 814-820 ◽

Cited By ~ 1

Author(s):

J A Bergeron ◽

J M DiNovo ◽

A F Razzano ◽

W J Dodds

Keyword(s):

Whole Blood ◽

Glass Bead ◽

Factor Xii ◽

Platelet Factor ◽

Serotonin Content ◽

Whole Blood Assay ◽

Vascular Integrity ◽

Blood Assay ◽

Standard Contact ◽

Recalcification Time

SummaryThe previously described native whole blood assay for materials in solution or suspension has been adapted to materials in a bead column configuration. These experiments showed that the glass itself accounts for little or none of the high blood-reactivity observed with conventional glass bead columns. Columns composed solely of soft glass that was “cleaned” by heat treatment (500-595° C 18 hr, electric oven) were benign toward flowing native whole blood for all variables measured (platelet count and platelet-free plasma [C14]-serotonin content, platelet factor 3 and factor XII activities, and recalcification time) with the standard contact protocol. In addition, the effluent successfully maintained perfusion of the isolated kidney, a measure of the ability of platelets to support vascular integrity. Prolonged (30 min) normothermic contact with titrated whole blood increased the subsequent reactivity of initially clean glass toward whole blood albeit to a level much less than that of conventional glass bead columns.

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