Effect of filgrastim (recombinant human granulocyte colony stimulating factor) on IgE responses in human asthma: a case study.

RATIONALE: The role of peripheral blood progenitor cell mobilization on Immunoglobulin E (IgE) responses has not been studied. METHODS: Distributions of blood lymphocytes (CD4+, CD8+, CD8+CD60+, CD19+, CD23+, CD16/56+, CD25, CD45RA+, CD45RO+, CD34+), and levels of serum immunoglobulins (IgM, IgG, IgA, IgE) were studied in an allergic asthmatic serum IgE+ (181IU/mL) adult (m/45 y/o) donor undergoing routine stem cell mobilization protocol (American Society of Hematology) before (day-30), during (day 4), and after (1 wk post last dose) filgrastim (subcutaneous, 480 mcg, 2qd) treatment (flow cytometry, nephelometry, UniCAP Total IgE Fluoro enzyme immunoassay). RESULTS: On day 4 of filgrastim treatment, numbers of CD8+CD60+T cells and CD23+ blood cells dramatically increased (98% and 240% respectively) compared with pre treatment. In contrast on day 4 of treatment, serum IgE levels decreased (>50%) compared with pre treatment. CD8+CD60+T cells and CD23+ blood cells and serum IgE levels approached pre-treatment levels at 1 week post treatment. CONCLUSIONS: Filgrastim treatment transiently increases numbers of CD8+CD60+T and CD23+ expressing cells, which are known to regulate human IgE responses, while also transiently suppressing ongoing IgE responses. These results suggest that filgrastim affects IgE related responses, and may be useful in modulating allergic responses.

Two distinct T cell subsets, CD4+ and CD8+CD60+, and their cytokines are required for in vitro induction of human ragweed-specific memory IgE responses.

CD8(+)CD60(+) T cells (80-98% CD45RO(+); 20% CD23(+)) are significantly increased in the blood of serum IgE(+) ragweed-sensitized (RS) compared with serum IgE-nonatopic humans (p = 0.001). CD8(+)CD60(+) T cells of the RS patients produced IL-2, IL-4, IL-10, IL-12, IFN-alpha. and IFN-gamma, but not IL-6 or IL-13. When their PBMC were cultured with ragweed Ag (RA), peak IgE responses occurred on day 10; none was induced with non-cross-reacting or without Ag; nonatopic PBMC did not respond to any stimulant. When either CD4(+) or CD8(+)CD60(+) T cells were depleted from RS PBMC before culture with RA, no IgE responses were induced. If purified CD4(+) T cells or low numbers of CD8(+)CD60(+) T cells were added back to the depleted PBMC, IgE responses were restored. However, higher numbers of CD8(+)CD60(+) T cells totally suppressed IgE responses. Total suppression also was obtained when RS PBMC were cultured with RA and either anti-IL-2, IL-4, IL-10, IL-12, IFN-gamma (all concentrations), or IFN-alpha (low concentrations), but not anti-IL-6 or IL-13. Higher concentrations of anti-IFN-alpha potentiated IgE responses.

Immune responses in autoimmune hepatitis: effect of prednisone and azathioprine treatment: case report.

The role of the immune response in autoimmune hepatitis has not been studied before and after prednisone and azathioprine treatment. Distributions of blood lymphocytes (CD4+, CD8+, CD19+, CD23+, CD16/56+), levels of serum immunoglobulins (IgM, IgG, IgE, IgA) and cytokines (IFN-gamma, IL-4, IL-12, TNFalpha ) were studied in a child (f/14 y/o) with autoimmune hepatitis before and after prednisone (20 mg/d) and azathioprine (50 mg/d) treatment (nephelometry, UniCAP Total IgE Fluoroenzymeimmunoassay, flow cytometry, cytokine ELISA). Patient was studied for 0-2.5 yrs; treatment was initiated 12 weeks post diagnosis. Numbers of CD4+ T cells increased (50%), while CD19+ and CD23+ cells decreased (>50%) post treatment; other lymphocyte subsets were unaffected by treatment. Serum IgG and IgE levels decreased (>50%) after treatment; serum IgM and IgA were within normal range and were not affected by treatment High levels of IFN-gamma (5-23 pg/ml) were initially detected in serum, which decreased after treatment (<0.1 pg/ml). Furthermore, low levels of IL-4 (0.2 pg/mL) were detected before treatment, which were not detected after treatment (<0.1 pg/ml). In contrast, before treatment, IL-12 and TNFalpha were not detected in serum; however after treatment the levels of IL-12 and TNFalpha dramatically increased. Prednisone and azathioprine treatment decreased total serum IgG, IgE, IFN-gamma and IL-4 levels, and blood CD19+ and CD23+ cells; however serum IL-12, TNFalpha and blood CD4+ T cells increased with treatment. Understanding immunomodulation in autoimmune hepatitis will provide better insight and mechanisms of this disease and may tailor more effective therapeutic intervention.

IgE, CD8(+)CD60+ T cells and IFN-alpha in human immunity to parvovirus B19 in selective IgA deficiency.

Although IgE is implicated in viral immunity, its role in parvovirus B19 immunity and its relationship to other immunological states has not been studied. Total serum immunoglobulin levels, IgG and IgE anti-parvovirus B19, blood lymphocyte numbers, and epsilon and cytokine specific mRNA were determined in pediatric patients with normal serum IgA levels (IgA+) and selective IgA deficiency (IgA-) on days 0 (initial diagnosis) and 14, and 3 years after recovery (nephelometry, Western blot test, flow cytometry, reverse transcriptase-polymerase chain reaction). We found that both patients had serum IgM, IgG, IgE, and IgA levels within normal ranges on day 0 to 3 years, excluding IgG(1) and IgA in the IgA- patient, which were elevated and negative, respectively, and IgE in the IgA+ patient, which was elevated (>100 IU/ml). The serum IgA+ and IgA- patients made IgE (and IgG) anti-parvovirus B19 at all time points. Excluding CD8(+)CD60+ T cells, determinations of T, B, and NK lymphocyte subsets always were within normal ranges. In both patients, CD8(+)CD60+ T-cell numbers were within normal ranges on day 0, but dramatically increased on day 14 (more than fivefold). At 3 years, they had returned to normal in the IgA+ patient, but remained high in the IgA- patient. On day 0 to 3 years, peripheral blood mononuclear cells of both patients expressed epsilon- and interferon (IFN)-alpha-specific mRNA. On day 0, the IgA+ patient expressed interleukin (IL)-4 and IL-10, but not IL-2, IFN-gamma, or IL-6 mRNA; the IgA- patient expressed IL-6 and IL-10 mRNA, but not IL-4, IL-2, or IFN-gamma mRNA. At 3 years, the IgA+ patient expressed mRNA for all cytokines, but the IgA- patient did not express mRNA for any of these cytokines. Our results suggest that IgE is important in parvovirus B19 immunity, and that IFN-alpha and CD8(+)CD60+ T cells may regulate IgE memory responses and isotype switching.

Immune response profiles after caterpillar exposure: a case report.

RATIONALE: The role of the immune response to caterpillar exposure is not well described. This case study is the first to report a patient who presented with an allergic reaction after exposure to the larvae of the sycamore tussock moth, Halysidota harrisii Walsh, 1864. METHODS: Blood was collected from an allergic asthmatic adult (m/42 y/o) at 2 hrs - 2 wks after contact urticaria with associated dyspnea after exposure to the larvae of the sycamore tussock moth, Halysidota harrisii Walsh, 1864. Distributions of blood lymphocytes (CD4(+), CD8(+), CD8(+)CD60(+), CD19(+), CD23(+), CD16/56(+), CD25, CD45RA(+), CD45RO(+)), monocytes (CD1d(+)), levels of serum immunoglobulins (IgM, IgG, IgA, IgE), and cytokines (IFN-γ, IL-4, TNF-α) were studied (flow cytometry, nephelometry, UniCAP Total IgE Fluoroenzymeimmunoassay, cytokine ELISA, clinical toxicology). RESULTS: Numbers of CD4(+) T cells, CD25(+) cells, CD19(+) B cells, and CD1d(+) monocytes decreased (22, 27, 33, 20%, respectively) one week post reaction, CD45RA(+) naïve T cells decreased at 36 hours (21%),while CD8(+)CD60(+) T cells and CD23(+) cells decreased 48 hrs (33, 74%, respectively) post reaction. In contrast, numbers of CD16/56(+) NK precursor cells increased (60%) 12 hrs, then decreased (65%) 48 hrs post reaction; other lymphocyte subsets were unaffected. Serum IgM, IgG and IgA were within normal range; however, serum IgE demonstrated a bimodal elevation at 2 hrs (15%) and one week post reaction. Levels of IFN-γ, IL-4, and TNF-α were not detected in serum pre-exposure (<1.0-4.0 pg/mL). However, high levels of IFN-γ (187-319 pg/mL) and TNF-α (549-749 pg/mL) were detected in serum 24-36 hrs and 3.5-24 hrs post reaction, respectively. In contrast, levels of IL-4 were undetected (<1.0 pg/mL) in serum at all time points. CONCLUSIONS: Exposure to the larvae of the sycamore tussock moth, Halysidota harrisii Walsh, 1864 may result in increased cytokine levels and blood CD16/56(+) NK precursor cells.

Green tea (Camelia sinensis) suppresses B cell production of IgE without inducing apoptosis.

Green tea (Camelia sinensis) is known to possess biological properties that are antioxidative and antimutagenic. Recent studies demonstrated beneficial effects of green tea in inflammatory allergy. However, the effect of green tea on anti-allergic activity/IgE responses in vitro has not been studied. U266 myeloma cells (2 x 10(6)/ml), which secrete IgE, were cultured for 0-72 hr with or without green tea extract (1-300 ng/ml), and IgE levels in the supernatants were determined (24-72 hr) by ELISA. The effects of green tea extract on U266 cell numbers, viability, and apoptosis were studied by flow cytometry. High levels of IgE produced by U266 cells were observed at 24, 48, and 72 hr (1.3 +/- 0.3 x 10(3), 1.7 +/- 0.3 x 10(3), 2.8 +/- 0.4 x 10(3) IU/ml, respectively). Addition of green tea extract either as (a) a single dose, or (b) repeated daily doses, suppressed IgE production with increasing suppression over time (up to 90%; p <0.05); the suppression was dose-dependent with the highest concentrations resulting in the greatest suppression. The suppression of IgE production by green tea extract was not mediated by apoptosis or cell death. This study demonstrates that green tea extract has immunoregulatory effects on human IgE responses in vitro.

IgE anti-varicella zoster virus and other immune responses before, during, and after shingles.

Blood lymphocyte distributions, serum immunoglobulin and cytokine levels, and serum IgE and IgG anti-varicella zoster virus (VZV) levels were measured in an atopic girl (age 15 yr) who developed shingles 10 yr after infection with chicken pox. Before, during, and 5 months after the shingles episode, the child's distributions of blood lymphocytes (excluding CD23+) and serum immunoglobulin levels (excluding IgE) were within the normal ranges. Her blood level of CD23+ lymphocytes decreased >50% during the shingles episode and remained low thereafter. Her serum level of IgE was elevated before and during shingles (154 and 168 IU/ml, respectively), but was reduced after recovery from shingles (<100 IU/ml). Before, during, and after shingles, her serum contained IgE and IgG anti-VZV antibodies. Before, during, and after shingles, low levels of IFN-gamma were detected in serum, but neither IL-1beta nor IL-4 were detected. Before shingles, low levels of IL-10 were detected in serum; during shingles, the serum level of IL-10 was increased 30-fold; it subsequently diminished at 5 mo after shingles. The role of IgE in immunity against varicella zoster virus (VZV) has not previously been studied. Our observations in this patient suggest that immunomodulation of IgE and accessory proteins may play a role in VZV pathogenesis.

Detection of IgE anti-parvovirus B19 and increased CD23+ B cells in parvovirus B19 infection: relation to Th2 cytokines.

The immune profile of a parvovirus B19-infected patient (male, 8 years old) was studied on day 0 (initial presentation) and on days 14 and 210 post symptom presentation (psp). Before infection, the patient was skin test positive to various allergens, including ragweed and tree and grass pollens, and had a serum IgE level of 150 IU/mL. On day 0, the patient was diagnosed as parvovirus B19 infected, as judged by the presence of IgG anti-parvovirus Abs in serum (EIA) and presentation of "slap cheek" rash. The patient's serum IgE level increased from 150 IU/mL before infection to 256 IU/mL on day 0, was 233 IU/mL on day 14, and returned to preinfection levels on day 210. In contrast, there was little change in the levels of serum IgM, IgG, or IgA (nephelometry). IgE anti-parvovirus B19 protein (VP-N) was detected in serum (Western blot) on days 0, 14, and 210, despite the decrease in total IgE on day 210. Although there was no increase in total numbers of blood CD23+ B cells on day 0, by day 14 the numbers of these cells increased dramatically (93%), remaining high on day 210. In contrast, there were virtually no changes in total numbers of CD4+ and CD8+ T cells or CD16/56+ NK precursor cells on days 0-210. On day 0, when IgG and IgE anti-parvovirus were detected in serum, patient's peripheral blood mononuclear cells (PBMC) expressed mRNA for the Th2 cytokines IL-4 and IL-10, but not for the Th1 cytokines IFN-gamma or IL-2. However, by day 14 psp, PBMC expressed mRNA for the Th1 cytokines IFN-gamma and IL-2, as well as for IL-4 and IL-10. This is the first demonstration of the existence of IgE anti-parvovirus B19 Ab. The presence of IgE anti-parvovirus B19 Ab in serum on day 0 and its persistence in serum 7 months psp suggests that IgE anti-parvovirus may be useful in prognosis of parvovirus B19 infection. Our results reinforce the idea that IgE, in general, may play a major role in anti-viral immunity, perhaps in conjunction with CD23+ cells. The results further suggest that clearance of this infection is accompanied by a switch to Th1 cytokines.

Effect of minocycline and doxycycline on IgE responses.

BACKGROUND: We have recently found that the tetracycline minocycline suppresses inflammatory responses in serum immunoglobulin (Ig)E-positive asthmatic patients, and that IgE levels can decrease in these patients. The mechanism by which minocycline suppresses these responses is unknown. OBJECTIVE: We have now investigated the ability of the tetracyclines, minocycline and doxycycline, to regulate IgE responses of peripheral blood mononuclear cells (PBMC) obtained from serum IgE-positive asthmatic patients. METHODS: The distributions of CD3+, CD4+, CD8+, and CD19+ lymphocytes in peripheral blood of serum IgE-positive asthmatic patients and IgE-negative nonasthmatic controls, and cytokine-specific mRNA expression by their PBMC were determined by flow cytometry (reverse transcriptase-polymerase chain reaction). Serum Ig levels also were determined (nephelometry, fluoroenzymeimmunoassay, enzyme-linked immunoadsorbent assay; n = 7/group). PBMC (1.5 x 10(6)/mL) were cultured with anti-CD40 monoclonal antibody and recombinant human interleukin-4 in the presence/absence of minocycline or doxycycline (0.1 to 10 microg/mL), and IgE levels in supernatants determined on days 0, 3, and 10 (enzyme-linked immunoadsorbent assay). RESULTS: Asthmatic and nonasthmatic subjects had similar numbers of blood CD4+ T cells (779/mm3 +/- 73 and 766 +/- 115, respectively) and CD19+ B-cells (239/mm3 +/- 35 and 379 +/- 95, respectively); however, CD8+ T cell numbers were decreased in asthmatic compared with nonasthmatic subjects (378/mm3 +/- 66 and 568 +/- 53, respectively; P = 0.045). High IgE levels were detected in supernatants of asthmatic PBMC on day 10 (28 ng/mL +/- 12), whereas control IgE levels did not change (<2.5 ng/mL). When either minocycline or doxycycline was included in culture, IgE production by asthmatic PBMC was strongly suppressed in dose-dependent fashion on day 10 (>80% with 10 microg/mL); control IgE did not change (<2.5 ng/mL). CONCLUSIONS: The results are consistent with the idea that the therapeutic benefits obtained by asthmatic patients from minocycline may, in part, result from IgE suppression.