• drug sensitivity
• granzyme a
• granzyme b
• granzyme k
• immune infiltration
• immunotherapy
• pan-cancer
• perforin 1
• tumor expression levels

• Autoimmune
• Autoinflammatory
• EBV
• Infections
• Inflammatory
• MAS
• Malignancies
• Rheumatic diseases
• Secondary
• pHLH
• sHLH

• Humans
• Lymphohistiocytosis, Hemophagocytic/genetics
• Lymphohistiocytosis, Hemophagocytic/immunology
• Lymphohistiocytosis, Hemophagocytic/therapy
• Cytokine Release Syndrome/genetics
• Cytokine Release Syndrome/immunology
• Cytokine Release Syndrome/etiology
• Genetic Predisposition to Disease

• Breast Cancer
• Drug Resistance
• Lysosomes
• Systems Biology Approach

• Classification
• HER2 positive breast cancer
• Immunogenomic profiling
• Machine learning
• Tumor immunity

• clinical guidelines
• cytolytic activity
• genes
• prognosis
• skin cutaneous melanoma
• therapies

• Wellcome Trust
• Wellcome
• European Research Council

• Granzyme B
• Perforin
• Staphylococcus enterotoxin C2

• Ankylosing spondylitis
• Bioinformatics analysis
• Crucial genes
• Immune infiltration

• Endometriosis
• Menstrual effluent
• Perforin
• T cells

• Multiple primary malignancies
• clinical characteristics
• lung cancer
• prognosis

• Aged
• Carcinoma, Squamous Cell/diagnosis
• Carcinoma, Squamous Cell/pathology
• Female
• Humans
• Incidence
• Lung Neoplasms/diagnosis
• Lung Neoplasms/pathology
• Male
• Middle Aged
• Neoplasms, Multiple Primary/diagnosis
• Neoplasms, Multiple Primary/pathology
• Neoplasms, Second Primary/diagnosis
• Neoplasms, Second Primary/pathology
• Prognosis
• Risk Factors

To determine whether adult cases of Chronic Lymphocytic Inflammation with Pontine Perivascular Enhancement Responsive to Steroids (CLIPPERS) may be related to familial hemophagocytic lymphohistiocytosis (HLH) causes, we have screened patients with adult-onset CLIPPERS for mutations in primary HLH-associated genes.

In our cohort of 36 patients fulfilling the criteria for probable or definite CLIPPERS according to the CLIPPERS-2017 criteria, we conducted a first study on 12 patients who consented to genetic testing. In these 12 patients, systemic HLH criteria were searched, and genetic analysis of 8 genes involved in primary HLH was performed.

Four definite and 8 probable CLIPPERS were enrolled (n = 12). Mutations involved in HLH were identified in 2 definite and 2 probable CLIPPERS (4/12). Three of them had biallelic PRF1 mutations with reduced perforin expression in natural killer cells. The remaining patient had biallelic UNC13D mutations with cytotoxic lymphocyte impaired degranulation. None of the mutated patients reached the criteria for systemic HLH. During follow-up, 3 of them displayed atypical findings for CLIPPERS, including emergence of systemic non-Hodgkin lymphoma (1/3) and confluent gadolinium-enhancing lesions on brain MRI (3/3).

In our patients presenting with adult-onset CLIPPERS, one-third have HLH gene mutations. This genetic treatable condition should be searched in patients with CLIPPERS, especially in those presenting with atypical findings.

• Actin cytoskeleton
• Cancer
• Exosomes
• Granzymes
• Immune evasion
• Immunological synapse
• Lysosomes
• Perforin
• Vesicle trafficking

• Actin Cytoskeleton/immunology
• Actin Cytoskeleton/pathology
• Actins/immunology
• Animals
• Humans
• Immunological Synapses/immunology
• Immunological Synapses/pathology
• Neoplasm Proteins/immunology
• T-Lymphocytes, Cytotoxic/immunology
• T-Lymphocytes, Cytotoxic/pathology
• Tumor Escape

• A91V
• A91V perforin

• Lymphoma
• bone marrow neoplasms
• cytogenetics
• diffuse
• large B-cell
• prognosis

Experimental models have demonstrated that immune surveillance by cytotoxic lymphocytes can protect from spontaneous neoplasms and cancer. In humans, defective lymphocyte cytotoxicity is associated with the development of hemophagocytic lymphohistiocytosis, a hyperinflammatory syndrome. However, to the best of the authors’ knowledge, the degree to which human lymphocyte cytotoxicity protects from cancer remains unclear. In the current study, the authors examined the risk of lymphoma attributable to haploinsufficiency in a gene required for lymphocyte cytotoxicity.

The authors exploited a founder effect of an UNC13D inversion, which abolishes Munc13‐4 expression and causes hemophagocytic lymphohistiocytosis in an autosomal recessive manner. Within 2 epidemiological screening programs in northern Sweden, an area demonstrating a founder effect of this specific UNC13D mutation, all individuals with a diagnosis of lymphoma (487 patients) and matched controls (1844 controls) were assessed using polymerase chain reaction for carrier status.

Among 487 individuals with lymphoma, 15 (3.1%) were heterozygous carriers of the UNC13D inversion, compared with 18 controls (1.0%) (odds ratio, 3.0; P = .002). It is interesting to note that a higher risk of lymphoma was attributed to female carriers (odds ratio, 3.7; P = .004).

Establishing a high regional prevalence of the UNC13D inversion, the authors have reported an overrepresentation of this mutation in individuals with lymphoma. Therefore, the results of the current study indicate that haploinsufficiency of a gene required for lymphocyte cytotoxicity can predispose patients to lymphoma, suggesting the importance of cytotoxic lymphocyte‐mediated surveillance of cancer. Furthermore, the results of the current study suggest that female carriers are more susceptible to lymphoma.

In the current study, the authors examine the risk of lymphoma attributable to an inversion disrupting UNC13D, a gene associated with familial hemophagocytic lymphohistiocytosis. The results demonstrate that haploinsufficiency of this gene, which is required for normal lymphocyte cytotoxicity, may predispose patients to lymphoma, signifying the importance of cytotoxic lymphocyte‐mediated surveillance of cancer development.

• cancer
• hemophagocytic lymphohistiocytosis
• immune surveillance
• lymphocyte cytotoxicity
• lymphoma

• cancer predisposition
• childhood leukemia
• down syndrome
• immune activation
• immune editing
• inflammation
• microbiome
• primary immunodeficiency

• Age Factors
• Animals
• Disease Susceptibility/immunology
• Environment
• Gene-Environment Interaction
• Genetic Predisposition to Disease
• Hematologic Neoplasms/epidemiology
• Hematologic Neoplasms/etiology
• Humans
• Immunologic Deficiency Syndromes/complications
• Immunologic Deficiency Syndromes/epidemiology
• Mutation
• Neoplasms/epidemiology
• Neoplasms/etiology

• Apoptosis
• Cancer immunotherapy
• Cell death
• Ferroptosis
• Granzymes
• Immunosurveillance
• Necroptosis
• Pyroptosis
• T cell

• Animals
• Cell Death/immunology
• Humans
• Neoplasms/immunology
• Neoplasms/pathology
• Neoplasms/therapy
• T-Lymphocytes/immunology

• K99 CA188679 NCI NIH HHS
• R00 CA188679 NCI NIH HHS

The immune system plays a significant role in urothelial bladder cancer (UBC) progression, with CD8⁺ T cells being capable to directly kill tumor cells using perforin and granzymes. However, tumors avoid immune recognition by escape mechanisms. In this study, we aim to demonstrate tumor immune escape mechanisms that suppress CD8⁺ T cells cytotoxicity. 42 patients diagnosed with UBC were recruited. CD8⁺ T cells from peripheral blood (PB), sentinel nodes (SN), and tumor were analyzed in steady state and in vitro-stimulated conditions by flow cytometry, RT-qPCR, and ELISA. Mass spectrometry (MS) was used for identification of proteins from UBC cell line culture supernatants. Perforin was surprisingly found to be low in CD8⁺ T cells from SN, marked by 1.8-fold decrease of PRF1 expression, with maintained expression of granzyme B. The majority of perforin-deficient CD8⁺ T cells are effector memory T (T_EM) cells with exhausted Tc2 cell phenotype, judged by the presence of PD-1 and GATA-3. Consequently, perforin-deficient CD8⁺ T cells from SN are low in T-bet expression. Supernatant from muscle invasive UBC induces perforin deficiency, a mechanism identified by MS where ICAM-1 and TGFβ2 signaling were causatively validated to decrease perforin expression in vitro. Thus, we demonstrate a novel tumor escape suppressing perforin expression in CD8⁺ T cells mediated by ICAM-1 and TGFβ2, which can be targeted in combination for cancer immunotherapy.

• Immune homeostasis
• immunotherapy
• melanoma

• Acute lymphoblastic leukemia
• Pediatric
• Perforin
• Survival

• adaptive natural killer cells
• chronic fatigue syndrome
• cytotoxic T cells
• lymphocyte cytotoxicity
• myalgic encephalomyelitis
• natural killer cells

• Autoimmune inflammatory syndrome induced by adjuvants (ASIA)
• Hemophagocytic lymphohistiocytosis (HLH)
• Intravascular large B-cell lymphoma (IVLBCL)
• Macrophage activation syndrome (MAS)
• Silicone breast implant
• Siliconosis

CD8 T cell-mediated blood-brain barrier (BBB) disruption is dependent on the effector molecule perforin. Human perforin has extensive single nucleotide variants (SNVs), the significance of which is not fully understood. These SNVs can result in reduced, but not ablated, perforin activity or expression. However, complete loss of perforin expression or activity results in the lethal disease familial hemophagocytic lymphohistiocytosis type 2 (FHL 2). In this study, we address the hypothesis that a single perforin allele can alter the severity of BBB disruption in vivo using a well-established model of CNS vascular permeability in C57Bl/6 mice. The results of this study provide insight into the significance of perforin SNVs in the human population.

We isolated the effect a single perforin allele has on CNS vascular permeability through the use of perforin-heterozygous (perforin+/−) C57BL/6 mice in the peptide-induced fatal syndrome (PIFS) model of immune-mediated BBB disruption. Seven days following Theiler’s murine encephalomyelitis virus (TMEV) CNS infection, neuroinflammation and TMEV viral control were assessed through flow cytometric analysis and quantitative real-time PCR of the viral genome, respectively. Following immune-mediated BBB disruption, gadolinium-enhanced T1-weighted MRI, with 3D volumetric analysis, and confocal microscopy were used to define CNS vascular permeability. Finally, the open field behavior test was used to assess locomotor activity of mice following immune-mediated BBB disruption.

Perforin-null mice had negligible CNS vascular permeability. Perforin-WT mice have extensive CNS vascular permeability. Interestingly, perforin-heterozygous mice had an intermediate level of CNS vascular permeability as measured by both gadolinium-enhanced T1-weighted MRI and fibrinogen leakage in the brain parenchyma. Differences in BBB disruption were not a result of increased CNS immune infiltrate. Additionally, TMEV was controlled in a perforin dose-dependent manner. Furthermore, a single perforin allele is sufficient to induce locomotor deficit during immune-mediated BBB disruption.

Perforin modulates BBB disruption in a dose-dependent manner. This study demonstrates a potentially advantageous role for decreased perforin expression in reducing BBB disruption. This study also provides insight into the effect SNVs in a single perforin allele could have on functional deficit in neurological disease.

• BBB disruption
• CD8 T cells
• PIFS
• Perforin
• Perforin single nucleotide variants
• TMEV

• Hereditary cancer
• perforin
• protein misfolding

The incidence of multiple primary malignancies (MPM) has increased sharply in recent decades. However, the clinical characteristics and prognosis of MPM patients involving lung cancer were not fully elucidated. This retrospective study was designed to explore the clinical characteristics and prognosis of MPM patients involving lung cancer in the People’s Republic of China.

Of 5405 lung cancer cases diagnosed at the Guangdong Lung Cancer Institute between 2005 and 2013, we analyzed 185 patients (3.4 %) with MPM involving lung cancer.

Among 185 patients with MPM involving lung cancer, 10 (5.4 %)had three malignancies and 175 (94.6 %) had two malignancies. 10 patients with three malignancies were excluded from the analysis to avoid misunderstanding. Of 175 accompanying malignancies, 64 (36.6 %) were synchronous MPM patients and 111 (63.4 %) were metachronous MPM patients; 49 (28.0 %) were lung cancer first MPM patients and 126 (72.0 %) were other cancer first MPM patients. The most frequent accompanying malignancy was colon cancer (25/175), followed by rectal cancer (18/175), esophageal cancer (17/175), and thyroid cancer (13/175). Metachronous MPM patients showed significantly better overall survival (OS) than synchronous MPM, with a median OS of 72.8 (range 12.2–391.0) and 12.9 (range 0.8–86.3)months, respectively (P < 0.001). Cox regression analysis revealed that time of occurrence and stage were independent factors for OS.

Colorectal cancer, esophageal cancer, and thyroid cancer were the tumors that most frequently accompanying lung cancer. Metachronous MPM patients showed significantly better OS compared with synchronous MPM patients.

• Hodgkin lymphoma
• T-cell lymphoma
• anaplastic large cell lymphoma
• hemophagocytic syndrome
• macrophage activation syndrome

• Lung transplantation
• NK cells
• natural killer cells
• non-Hodgkin lymphoma
• perforin

Hemophagocytic lymphohistiocytosis (HLH) is a hyperinflammatory syndrome that develops as a primary (familial/hereditary) or secondary (non-familial/hereditary) disease characterized in the majority of the cases by hereditary or acquired impaired cytotoxic T-cell (CTL) and natural killer responses. The molecular mechanisms underlying impaired immune homeostasis have been clarified, particularly for primary diseases. Familial HLH (familial hemophagocytic lymphohistiocytosis type 2-5, Chediak-Higashi syndrome, Griscelli syndrome type 2, Hermansky-Pudlak syndrome type 2) develops due to a defect in lytic granule exocytosis, impairment of (signaling lymphocytic activation molecule)-associated protein, which plays a key role in CTL activity [e.g., X-linked lymphoproliferative syndrome (XLP) 1], or impairment of X-linked inhibitor of apoptosis, a potent regulator of lymphocyte homeostasis (e.g., XLP2). The development of primary HLH is often triggered by infections, but not in all. Secondary HLH develops in association with infection, autoimmune diseases/rheumatological conditions and malignancy. The molecular mechanisms involved in secondary HLH cases remain unknown and the pathophysiology is not the same as primary HLH. For either primary or secondary HLH cases, immunosuppressive therapy should be given to control the hypercytokinemia with steroids, cyclosporine A, or intravenous immune globulin, and if primary HLH is diagnosed, immunochemotherapy with a regimen containing etoposide or anti-thymocyte globulin should be started. Thereafter, allogeneic hematopoietic stem-cell transplantation is recommended for primary HLH or secondary refractory disease (especially EBV-HLH).

• Alemtuzumab
• Anti-thymocyte globulin
• Cyclosporine A
• Epstein-Barr virus
• Etoposide
• Hematopoietic stem-cell transplantation
• Hemophagocytic lymphohistiocytosis
• Hereditary diseases
• Immunochemotherapy
• Intravenous immunoglobulin
• Molecular diagnosis
• Rituximab
• Steroids

• Auto-Immune-Related Hematocytopenia
• IFN-γ
• IL-17A
• eosinophile
• granzyme
• perforin

The human immune system depends on the activity of cytotoxic T lymphocytes (CTL), natural killer (NK) cells, and NKT cells in order to fight off a viral infection. Understanding the molecular mechanisms during this process and the role of individual proteins was greatly improved by the study of familial hemophagocytic lymphohistiocytosis (FHL). Since 1999, genetic sequencing is the gold standard to classify patients into different subgroups of FHL. The diagnosis, once based on a clinical constellation of abnormalities, is now strongly supported by the results of a functional flow-cytometry screening, which directs the genetic study. A few additional congenital immune deficiencies can also cause a resembling or even identical clinical picture to FHL. As in many other rare human disorders, the collection and analysis of a relatively large number of cases in registries is crucial to draw a complete picture of the disease. The conduction of prospective therapeutic trials allows investigators to increase the awareness of the disease and to speed up the diagnostic process, but also provides important functional and genetic confirmations. Children with confirmed diagnosis may undergo hematopoietic stem cell transplantation, which is the only cure known to date. Moreover, detailed characterization of these rare patients helped to understand the function of individual proteins within the exocytic machinery of CTL, NK, and NKT cells. Moreover, identification of these genotypes also provides valuable information on variant phenotypes, other than FHL, associated with biallelic and monoallelic mutations in the FHL-related genes. In this review, we describe how detailed characterization of patients with genetic hemophagocytic lymphohistiocytosis has resulted in improvement in knowledge regarding contribution of individual proteins to the functional machinery of cytotoxic T- and NK-cells. The review also details how identification of these genotypes has provided valuable information on variant phenotypes.

• cellular cytotoxicity
• hemophagocytosis
• mutation analysis
• natural killer

• autophagy
• breast cancer
• colorectal cancer
• endoplasmic stress
• immunogenic cell death
• tumor-infiltrating lymphocytes

• Animals
• Diabetes Mellitus, Type 1/genetics
• Diabetes Mellitus, Type 1/immunology
• Graft Rejection/genetics
• Graft Rejection/immunology
• Humans
• Immune System Diseases/genetics
• Immune System Diseases/immunology
• Models, Molecular
• Perforin/chemistry
• Perforin/genetics
• Perforin/physiology

Congenital perforin deficiency is considered a rare cause of human immunopathology and immune dysregulation, and classically presents as a fatal illness early in infancy. However, we propose that a group of related disorders in which killer lymphocytes deliver only partially active perforin or a reduced quantum of wild-type perforin to the immune synapse should be considered part of an extended syndrome with overlapping but more variable clinical features. Apart from the many rare mutations scattered over the coding sequences, up to 10% of Caucasians carry the severely hypomorphic PRF1 allele C272 > T (leading to A91V mutation) and the overall prevalence of the homozygous state for A91V is around 1 in 600 individuals. We therefore postulate that the partial loss of perforin function and its clinical consequences may be more common then currently suspected. An acute clinical presentation is infrequent in A91V heterozygous individuals, but we postulate that the partial loss of perforin function may potentially be manifested in childhood or early adulthood as “idiopathic” inflammatory disease, or through increased cancer susceptibility – either hematological malignancy or multiple, independent primary cancers. We suggest the new term “perforinopathy” to signify the common functional endpoints of all the known consequences of perforin deficiency and failure to deliver fully functional perforin.

• FHL
• NK cell
• granzyme
• immune deficiency
• perforin
• perforinopathy
• protein misfolding