Systemic evaluation and localization of resistin expression in normal human tissues by a newly developed monoclonal antibody

Resistin and resistin-like molecules are pleiotropic cytokines that are involved in inflammatory diseases. Our previous work suggested that resistin has the potential to be used as a biomarker and therapeutic target for human pulmonary arterial hypertension. However, data are limited on the distribution of resistin in healthy human organs. In this study, we used our newly developed anti-human resistin (hResistin) antibody to immunohistochemically detect the expression, localization, and intracellular/extracellular compartmentalization of hResistin in a full human tissue panel from healthy individuals. The potential cross reactivity of this monoclonal anti-hResistin IgG1 with normal human tissues also was verified. Results showed that hResistin is broadly distributed and principally localized in the cytoplasmic granules of macrophages scattered in the interstitium of most human tissues. Bone marrow hematopoietic precursor cells also exhibited hResistin signals in their cytoplasmic granules. Additionally, hResistin labeling was observed in the cytoplasm of nervous system cells. Notably, the cytokine activity of hResistin was illustrated by positively stained extracellular material in most human tissues. These data indicate that our generated antibody binds to the secreted hResistin and support its potential use for immunotherapy to reduce circulating hResistin levels in human disease. Our findings comprehensively document the basal expression patterns of hResistin protein in normal human tissues, suggest a critical role of this cytokine in normal and pathophysiologic inflammatory processes, and offer key insights for using our antibody in future pharmacokinetic studies and immunotherapeutic strategies.


Negative Control Material
Human hypercalcemia of malignancy peptide, amino acid residues 1-34, UV-resin spot slides PTHrP No staining of negative control PTHrP 1-34 protein spots with either concentration of hResistin IgG or HuIgG1 or in the assay control slide. Incomplete washing of slide 1 in runs 2 and 3; did not preclude interpretation.
General Comments: All tissue specimens judged adequate for interpretation unless otherwise specified. Background staining due to incompletely quenched endogenous myeloperoxidase or endogenous/exogenous pigments described for individual tissues.
Nuclear staining of multiple cell types was observed in the majority of tissues. This staining was sometimes quite intense in slide 1 and reduced in intensity and frequency in slide 2. Nuclear staining was judged nonspecific and hampered but did not preclude interpretation.
Slides were numbered according to the following scheme: Slide 1 (hResistin IgG, 20 µg/mL), Slide 2 (hResistin IgG, 5 µg/mL), Slide 3 (HuIgG1, 20 µg/mL), Slide 4 (HuIgG1, 5 µg/mL), Slide 5 (Assay Control [omit primary antibody]), Slide 6 (Anti-β2-microglobulin [tissue staining control]). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type).

Adrenal
Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type).

Neg
Neg Neg Observed in interstitium/stroma particularly in perivascular areas. Staining often appeared as diffuse to finely punctate material and often lined up on extracellular matrix fibers. ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type).
Neg Neg Observed scattered in interstitium.
Cells/processes associated with peripheral nerves (cytoplasm) Neg Neg Includes myenteric plexus.
Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type).

Neg
Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type).

Neg
Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type).

Neg
Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). Other elements Neg Neg Neg Neg Neg ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type). 1-3+ (occas to freq) ± = equivocal, 1+ = weak, 2+ = moderate, 3+ = strong, 4+ = intense, Neg = Negative, Pos = Positive, M = Missing, NE = Not Evaluated, NS = Not Stained, freq = frequent, occas = occasional. Frequency modifiers were included to provide the approximate percentage staining of expected numbers of that cell type or tissue element at that location. The frequency of cells with staining was identified as follows: very rare (<1% of cells of a particular cell type); rare (1-5% of cells of a particular cell type); rare to occasional (>5-25% of cells of a particular cell type); occasional (>25-50% of cells of a particular type); occasional to frequent (>50-75% of cells of a particular cell type); frequent (>75-100% of cells of a particular cell type).