Pathophysiological implication of reversed CT halo sign in invasive pulmonary mucormycosis: a rare case report
© Okubo et al.; licensee BioMed Central Ltd. 2013
Received: 18 January 2013
Accepted: 8 May 2013
Published: 17 May 2013
It has been accepted that reversed halo sign (RHS) appeared on a computed tomography (CT) image in immunocompromised patients indicates an invasive fungal infection, but its pathophysiology remains obscure as to what this image implies. Therefore, the present report describes detailed radiological and histopathological findings of a case of invasive pulmonary mucormycosis (IPM) presenting RHS with comparison to those from a lesion of discrete nodule caused by invasive pulmonary aspergillosis (IPA), and discusses the pathophysiological implications of this characteristic image.
RHS had been clinically noted at the time of recovering of bone marrow function of a 64-year-old Japanese man who had chemotherapy for his acute lymphoblastic leukemia. Histological examination of the surgically removed lung revealed a lesion of IPM. This was composed of coagulation necrosis of septa at the center of lesion with preservation of air content which was encompassed outer rim comprising triplet structure; liquefaction, consolidation, and organization from the inner to the outer layer. In addition, Micro-CT examination confirmed reticular structure and monotonous high density at the central coagulation necrosis preserving air content and surrounding consolidation, and organization lesion of the IPM lesion.
Our investigations suggest that RHS might be understood as a kind of immune reconstitution syndrome and be the initial and prior status of air crescent sign.
The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/3480054198968132
KeywordsReversed halo sign Mucormycosis Aspergillosis Discrete nodule
Reversed halo sign (RHS) has been regarded as a sign on computed tomography (CT) image characterized by central ground-glass opacity (GGO) which is surrounded by a partial or complete rim of consolidation. Although this sign was initially described specifically for cryptogenic organizing pneumonia , it has also been associated with other diseases, including fungal infections [2, 3]. However, only a few studies have conducted a detailed pathophysiological investigation of this phenomenon. Therefore, the present report describes detailed radiological and histopathological findings of a case of invasive pulmonary mucormycosis (IPM) presenting RHS, with comparison to a discrete nodule (DN) [4, 5] caused by invasive pulmonary aspergillosis (IPA), and discusses the pathophysiological implications of this characteristic image.
Material and methods
Lung tissue from surgically removed the patient with IPM presenting RHS on a CT image was prepared for the present report. Additionally, lung tissue from autopsied patients with IPA was also employed to aid in a discussion of the pathophysiological implications of RHS. Namely, a representative sample of IPA was chosen among the lung tissues of 64 autopsies with IPA, which has been previously reported . The selection was based on the following two criteria: one is a patient who had an agranulocytosis due to severe bone marrow suppression and another is the formalin-fixed tissue from IPA indicating a discrete nodular coagulation necrosis with surrounding of oozing; DN, which has been generally accepted as the lesion mirrored as CT halo sign [4, 5]. In addition, written informed consent (in Japanese) was obtained from the patients’ family for publication of this study and any accompanying images. The case report was undertaken in compliance with the Helsinki Declaration.
Routine histopathological examination was conducted. Briefly, the specimens were fixed with 10% formalin, embedded in paraffin wax after dehydration, and cut into 4 μm-thick sections. These were then prepared and stained with Hematoxylin and Eosin double, Elastica van Gieson, and Grocott stains for light microscopic observation.
Image analyzing with micro-CT and 3D image reconstruction
The formalin-fixed tissues were dehydrated with 100% ethanol for 24 hours, dried for 3 hours at room temperature, and trimmed to a proper size. Scanning was carried out using a micro-CT scanner (InspeXio SMX-100CT, Shimadzu, Kyoto, Japan) with 45 kV tube voltage and 40 μA tube electric current. In the present study, the field of view was fixed at approximately six mm (XY plane) X five mm (Z plane) and the reconstruction matrix size was determined 512 × 512 pixels. After obtaining more than 400 two-dimensional (2 D) images of 12-μm thickness, a series of individual 2 D images were provided to a virtual three-dimensional (3 D) image . Namely, parallel 2 D images were stacked using a commercially available 3 D reconstruction program (VG studio software, Nihon Visual Science, Tokyo, Japan) to generate 3 D images of the tissue samples. In the present report, since representative tissue samples from patients with IPM or IPA was prepared one from each lung tissue, a statistical analysis was not used.
It has been accepted that RHS has appeared in CT images from immunocompromised patients indicates invasive fungal infection , including in patients with pulmonary mucormycosis . In particular, Busca A et al. described that RHS may be an early radiological sign of mucormycosis and emphasized the importance of recognizing this sign as a diagnostic strategy . However, detailed pathophysiology of the sign remains obscure. Therefore, we would like to discuss how RHS develops in comparison with the histopathological findings and micro-CT images of pulmonary lesions presented IPM or IPA. For our patient, the lesion presented RHS histologically comprised central coagulation necrosis remaining air content and ring-shaped peripheral consolidation or fibrosis, which was also confirmed by micro-CT images and their 3D reconstruction. The central GGO had been clinically noted during neutropenia prior to out-ward development of the outer rim, which was coincidently confirmed at the time of recovering of PNC. This fact may suggest that the recovery of impaired bone marrow function derived neutrophils at the periphery of coagulation necrosis, which might be induced by intravascular growth of invaded Mucor. Neutrophil provision might induce thin ring-shaped liquefaction between coagulation necrosis and adjacent unaltered parenchyma because there were no elastic fibers in the liquefaction, which was digested by neutrophil function. The liquefaction, logically understood as abscess was followed by the out-ward development of mononuclear cell infiltrate replacing alveolar space which as observed as the second layer of the outer rim of CT-RHS. This finding is generally accepted as a protective response against necrosis, but this was not developed inside the ring-shaped liquefaction because coagulation necrosis had no blood flow . Further protective response was noted as the third layer of the outer rim, which can be named as organization . The change is histologically defined as fibrosis entirely filling the alveolar space and well-remaining elastin in septa. Accordingly, pathophysiology of CT-RHS might be understood as a kind of immune reconstitution syndrome. In the case, the sign might mirror the recovering of impaired bone marrow functioning. On the other hand, it has been reported that recovery of neutrophil function in patients with IPA result in liquefaction and this process was recognized prior to the development of a CT-air crescent sign . In the present IPM case, the CT image on -11st day revealed air crescent-like small air slit between GGO and the outer rim, which might be reflected by the excretion of liquefaction layer. It suggests that RHS might be the initial and prior status of CT-air crescent sign.
The present study also shed some light on the difference in both histological and micro-CT images between IPM and IPA in patients with neutropenia. The lung tissue histologically presented DN comprised nodule of coagulation without air content, where mirrored the nodule of monotonous high density on the micro-CT image. Accordingly, a central monotonous high-density nodule and GGO may be understood as a characteristic sign of IPA and IPM in patient with neutropenia, respectively. The difference should be derived whether alveolar space is filled or not filled with exudation. However, cause of exudation filling alveolar space in case of IPA is unclear, although the difference in intensity of penetration can be suggested between Aspergillus and Mucor. Namely, Aspergillus usually shows a strong intensity of penetration in the lung regardless of a difference in compliance and/or elasticity for each component. Such strong invasiveness of Aspergillus may easily induce plasma exudation. In contrast, Mucor could penetrate though septal wall from lumen of capillary or small vascular, if exudation were seen in alveoli like IPA.
Meanwhile, we wish to go into insight of the usefulness of the molecular supplemental procedures on formalin-fixed and paraffin-embedded (FFPE) tissue sections. Whereas it has been largely accepted that microscopic findings of the hyphae (diameter, presence of septa, branching angle, and pigmentation) are important to determine causative agents in foci of invasive mold infections, it might be difficult to identify causative molds only by histopathological observation even that was made with the best effort. Histological identification might have a limitation itself . Therefore, the establishment of an auxiliary diagnostic method for use in routine pathological laboratories is required. Previous investigators emphasized the usefulness of molecular biology methods on FFPE tissue sections [11–13]. Since FFPE sections are widely employed in routine preparations for surgical pathology, molecular biology methods may be utilized in many medical institutes. Furthermore, an additional diagnostic procedure will not be required if the stored paraffin-embedded tissue could be utilized for considerable molecular methods. Although further careful investigations are required, an establishment of accurate auxiliary molecular biology method could surely contribute to improve an accuracy of conventional histopathological diagnosis of this seriously debilitating infectious disease.
Our study can suggest that reversed halo sign might be considered as a form of immune reconstitution phenomena, especially induced by recovering of bone marrow functioning, and be the initial and prior lesion of the air crescent sign.
Written informed consent was obtained from the patient for publication of this Case report and any accompanying images. A copy of the written consent is available for review by the Editor of this journal.
Reversed halo sign
Invasive pulmonary mucormycosis
Invasive pulmonary aspergillosis
Peripheral neutrophil count
Formalin-fixed and paraffin-embedded.
This work was supported by the Health Science Research Grants for Research on Emerging and Re-emerging Infectious Diseases (H22-Shinkou-Ippan-8 and H23-Shinkou-Ippan-018) from the Ministry of Health, Labor and Welfare of Japan, a grant from the Strategic Basis on Research Grounds for Non-governmental Schools at Heisei 20th, the Strategic Research Foundation Grant-aided Project for Private schools at Heisei 23rd, and KAKENHI to Yoichiro Okubo (#24790364) and Daisuke Sasai from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, Toho University project grant #23-19, 21, and 28 and #24-11, 16, 27, and 28, Yokohama Foundation for Advancement of Medical Science grant to YO and MS, Dr Yanase’s grant of Toho University Medical School to YO. Authors thank Toho University, Medical Media Center staff members for their excellent technical support as well as Yoshiro Yamamoto, Kayoko Shimodaira, and Zhi Yuzhu for their histopathological advice. In addition, authors thank Hirokazu Okochi and Eiji Otawa (Product Experts Group, Global Marketing Department, Analytical & Measuring Instruments Division, Shimadzu Corporation) for their technical support.
- Voloudaki AE, Bouros DE, Froudarakis ME, Datseris GE, Apostolaki EG, Gourtsoyiannis NC: Crescentic and ring-shaped opacities. CT features in two cases of bronchiolitis obliterans organizing pneumonia (BOOP). Acta Radiol. 1996, 37 (6): 889-892. 10.3109/02841859609175463.View ArticlePubMedGoogle Scholar
- Marchiori E, Zanetti G, Escuissato DL, Souza AS, Meirelles GSP, Fagundes J, Souza CA, Hochhegger B, Marom EM, Godoy MC: Reversed halo sign: high-resolution CT scan findings in 79 patients. Chest. 2012, 141 (5): 1260-1266. 10.1378/chest.11-1050.View ArticlePubMedGoogle Scholar
- Georgiadou SP, Sipsas NV, Marom EM, Kontoyiannis DP: The diagnostic value of halo and reversed halo signs for invasive mold infections in compromised hosts. Clin Infect Dis. 2011, 52 (9): 1144-1155. 10.1093/cid/cir122.PubMed CentralView ArticlePubMedGoogle Scholar
- Shibuya K, Ando T, Hasegawa C, Wakayama M, Hamatani S, Hatori T, Nagayama T, Nonaka H: Pathophysiology of pulmonary aspergillosis. J Infect Chemother. 2004, 10 (3): 138-145.View ArticlePubMedGoogle Scholar
- Greene R, Shibuya K, Ando T: Diagnosis. Aspergillus fumigatus and aspergillosis. Edited by: Latgé J-P, Steinbach William J. 2009, Washington D.C: ASM Press, 353-362.View ArticleGoogle Scholar
- Wetzler M, Dodge RK, Mrozek K, Carroll AJ, Tantravahi R, Block AW, Pettenati MJ, Le Beau MM, Frankel SR, Stewart CC: Prospective karyotype analysis in adult acute lymphoblastic leukemia: the cancer and leukemia Group B experience. Blood. 1999, 93 (11): 3983-3993.PubMedGoogle Scholar
- Kato A, Ohno N: Construction of three-dimensional tooth model by micro-computed tomography and application for data sharing. Clin Oral Investig. 2009, 13 (1): 43-46. 10.1007/s00784-008-0198-4.View ArticlePubMedGoogle Scholar
- Busca A, Limerutti G, Locatelli F, Barbui A, De Rosa FG, Falda M: The reversed halo sign as the initial radiographic sign of pulmonary zygomycosis. Infection. 2012, 40 (1): 77-80. 10.1007/s15010-011-0156-y.View ArticlePubMedGoogle Scholar
- Kumar V, Abbas AK, Aster JC, Robbins SL: Inflammation and repair. Robbins basic pathology. Edited by: Kumar V, Abbas AK, Aster JC, Robbins SL. 2013, Philadelphia: Elsevier/Saunders, 29-74. 9Google Scholar
- Travis WD, Colby TV, Koss MN, Rosado-de-Christenson ML, Müller NL, Talmadge E, King J: Atlas of nontumor pathology Non-Neoplastic Disorders of the Lower Respiratory Tract. Edited by: West King D. 2001, Washington DC: American Registry of Pathology, 592-636.Google Scholar
- Hofman V, Dhouibi A, Butori C, Padovani B, Gari-Toussaint M, Garcia-Hermoso D, Baumann M, Venissac N, Cathomas G, Hofman P: Usefulness of molecular biology performed with formaldehyde-fixed paraffin embedded tissue for the diagnosis of combined pulmonary invasive mucormycosis and aspergillosis in an immunocompromised patient. Diagn Pathol. 2010, 5: 1-10.1186/1746-1596-5-1.PubMed CentralView ArticlePubMedGoogle Scholar
- Shinozaki M, Okubo Y, Sasai D, Nakayama H, Murayama SY, Ide T, Wakayama M, Ishiwatari T, Tochigi N, Nemoto T: Development of a peptide nucleic acid probe to Trichosporon species and identification of trichosporonosis by use of in situ hybridization in formalin-fixed and paraffin-embedded (FFPE) sections. J Clin Microbiol. 2013, 51 (1): 295-298. 10.1128/JCM.02221-12.PubMed CentralView ArticlePubMedGoogle Scholar
- Okubo Y, Shinozaki M, Wakayama M, Nakayama H, Sasai D, Ishiwatari T, Nemoto T, Naobumi T, Shibuya K: Applied gene histopathology: identification of Fusarium species in FFPE tissue sections by in situ hybridization. Methods Mol Biol. 2013, 968: 141-147. 10.1007/978-1-62703-257-5_10.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.